Trispecific T cell engagers

Provided are trispecific T Cell Engagers or TSMAb's, antibodies that can simultaneously engage three different types of epitopes on the same target or on different targets. More specifically, the invention is directed to trispecific molecules that bind to DLL3, MUC17 or CLDN18.2 and activate CD (cluster of differentiation) molecules (e.g. CD3, CD28 and CD137). Also provided are methods of treating an ailment such as cancer using an antibody (or fragment) against DLL3, MUC17 or CLD18 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 and/or CD137.

1. 11. An anti-CD28, anti-CD3, and anti-DLL3 trispecific antibody, comprising: a first peptide chain comprising (1) an anti-CD28 single chain fragment variable region (scFv), (2) an anti-CD3 scFv and (3) one peptide chain of a fragment crystallizable region (Fc region); a second peptide chain comprising (1) one peptide chain of an anti-DLL3 Fab fragment and (2) another peptide chain of the Fc region; and a third peptide chain comprising another peptide chain of the anti-DLL3 Fab fragment, and wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 276 and the third peptide chain comprises SEQ ID NO: 302; or wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 291 and the third peptide chain comprises SEQ ID NO: 295; or wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 290 and the third peptide chain comprises SEQ ID NO: 298; or wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 276 and the third peptide chain comprises SEQ ID NO: 297.
2. 22. The trispecific antibody of claim 1, wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 276 and the third peptide chain comprises SEQ ID NO: 302.
3. 33. The trispecific antibody of claim 1, wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 291 and the third peptide chain comprises SEQ ID NO: 295.
4. 44. The trispecific antibody of claim 1, wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 290 and the third peptide chain comprises SEQ ID NO: 298.
5. 55. The trispecific antibody of claim 1, wherein the first peptide chain comprises SEQ ID NO: 481, the second peptide chain comprises SEQ ID NO: 276 and the third peptide chain comprises SEQ ID NO: 297.

RELATED APPLICATIONS
This application is a continuation application of U.S. patent application Ser. No. 17/164,696, filed on Feb. 1, 2021, which claims priority to U.S. provisional patent application Ser. No. 62/968,999 filed on Jan. 31, 2020, U.S. provisional patent application Ser. No. 62/981,048 filed on Feb. 25, 2020 and U.S. provisional patent application Ser. No. 62/991,070 filed on Mar. 17, 2020. The contents of the aforementioned applications are incorporated herein by reference.


REFERENCE TO SEQUENCE LISTING
The application contains a Sequence Listing which has been submitted electronically in .XML format and is hereby incorporated by reference in its entirety. Said .XML copy, created on Nov. 26, 2024, is named “2022-007 CONT.xml” and is 770,597 bytes in size. The sequence listing contained in this .XML file is part of the specification and is hereby incorporated by reference herein in its entirety.
FIELD OF INVENTION
This invention relates generally to cancer therapies, and more specifically, to novel compounds comprising anti-DLL3, CLDN18.2 and Muc17 antibodies or immunoreactive fragments thereof for the treatment of cancer.
BACKGROUND
Cancer is generally defined as a group of diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. In 2019, roughly 1.8 million people were diagnosed with cancer in the United States. Each year, an estimated 606,880 people will die from cancer in the United States. Lung and bronchus cancer is responsible for the most deaths. Colorectal cancer and pancreatic cancer are the second and third most common causes of cancer death respectively.
Cancer has been linked to several factors including smoking, obesity, poor diet, lack of physical activity and excessive consumption of alcohol. Other factors include certain infections, exposure to ionizing radiation and environmental pollutants. Certain cancers have been linked to infections such as Helicobacter pylori, hepatitis B, hepatitis C, human papillomavirus infection, Epstein-Barr virus and human immunodeficiency virus (HIV).
Conventional cancer treatments are directed at removing cancerous tissue and preventing it from spreading. Such treatment options include surgery, chemotherapy, radiation therapy, hormonal therapy, targeted therapy and palliative care. Treatments are usually pursued based on the type, location and grade of the cancer as well as the patient's health and preferences. These options have limitations. They can be ineffective, particularly when cancer has metastasized. Moreover, chemotherapy and radiation therapy have a range of side-effects related to cell toxicity.
Because cancer cells divide faster than most normal cells, they can be sensitive to chemotherapy drugs. However, chemotherapy drugs will also attack other cells in the body, especially fast-dividing cells such as blood cells and the cells lining the mouth, stomach, and intestines. Accordingly, there is a need for improved medications and methods of treating cancer that are more targeted and have less deleterious side effects.
A promising area for the development of treatments includes targeted therapies using antibodies. For example, the use of antibody drug conjugates can be used to target a drug toward a tumor. Immunoconjugates are antibodies conjugated joined) to a second molecule, usually a toxin, radioisotope or label. Immunoconjugates can provide for relatively high concentrations of drug within the tumor whereas systemic administration of unconjugated (i.e., untargeted) drug to achieve the same tumor concentration can lead to levels that are toxic to normal cells.
Another promising area is development of treatments that harness the immune system to attack and kill tumor cells. Checkpoint inhibitors, such as anti-CTLA4, anti-PD1 and anti-POL1 therapies have changed the way cancer is treated. Similarly, the direct activation of cytotoxic T cells by bispecific T cell engagers or CAR-T engineered T cells, has led to previously unseen cures in many types of cancers.
Delta-like ligand 3 (DLL3) is an inhibitory notch ligand that is expressed at relatively low levels in normal tissues. It is expressed at high levels in small cell lung cancer (SCLC) and other neuroendocrine tumors, thus presenting potential therapeutic target in cancer diagnosis and treatment.
Mucin 17, also referred to as MUC17, is a member of the mucin family that is composed of more than 20 members. Mucins are large, highly glycosylated membrane bound proteins. They generally function in mucosal areas to protect epithelial cells from their environment, as well as to regulate proliferation and survival of cells. MUC17 is expressed in pancreatic, appendiceal, and some colon cancers and thus is a target antigen for these cancers. Thus, MUC17 is a candidate for targeting of therapies such as antibody drug conjugates, T cell engagers, and CAR-T cells.
Claudin-18 (CLDN18) is a protein in humans that is encoded by the CLDN18 gene. It belongs to the group of claudins, a family of proteins that form components of tight cell junction strands in epithelial cells. Studies have demonstrated that Isoform 2 (Claudin 18.2 or CLDN18.2) is abundant in gastric tumors. It has exposed extracellular loops and is available for monoclonal antibody binding. These biological characteristics have led to the development of monoclonal antibodies against claudin 18.2, such as claudiximab (IMAB362).
CD3, CD28 and CD137 are receptors present on T-cells. T cells can be activated though CD3, CD28 and CD137, by antigen-presenting cells that utilize the activation signals MHC Class I and II, CD80 and CD86, and 4-1BBL, respectively. CD3 is part of the T cell receptor (TCR) and is the signaling component for the receptor. There are three CD3 subunits, epsilon, delta and gamma. Epsilon associates with both delta and zeta and together they are responsible for signaling. CD3 signaling is considered signal 1 that is required to activate T cells. The co-receptors, CD28 and CD137, are considered signal 2. Both signal 1 and signal 2 are required for full activation, proliferation and survival of T cells.
The present invention discloses trispecific T Cell Engagers. The trispecific and trispecific molecules can bind to DLL3, MUC17 and/or CLDN18.2 and activate CD (cluster of differentiation) molecules (e.g. CD3, CD28 and CD137). Also provided are methods of treating an ailment such as cancer using antibodies and antibody conjugates, pharmaceutical compositions thereof, and articles of manufacture.
SUMMARY
The inventions described and claimed herein have many attributes and embodiments including, but not limited to, those set forth or described or referenced in this brief summary. The inventions described and claimed herein are not limited to, or by, the features or embodiments identified in this summary, which is included for purposes of illustration only and not restriction.
An aspect of the invention is an antibody against DLL3. The antibody can be a fragment such as an antigen binding fragment (Fab) or a single chain variable fragment (Scfvs).
An aspect of the invention is an agonist antibody that activates CD3, CD28 and/or CD137.
An aspect of the invention is a bispecific or trispecific molecule that includes an antibody (or fragment) against DLL3 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a method of treating an ailment such as cancer using an antibody (or fragment) against DLL3 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a method of treating an ailment that uses two or more of the trispecific molecules described herein in combination with one another.
An aspect of the invention is a method of treating an ailment that uses one bispecific and one trispecific molecule described herein in combination with one another.
An aspect of the invention is a method of treating an ailment that uses two or more trispecific molecules described herein in combination with one another.
An aspect on the invention is a method of activating T-cell cytotoxicity against DLL3 expressing cells.
An aspect of the invention is a method of activating T-cell cytotoxicity using a bispecific molecule the includes an antibody (or fragment) against DLL3 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a humanized antibody which binds to human DLL3 protein comprising a heavy chain variable domain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% sequence identity to SEQ ID NO: 56-74 or 75.
The disclosed methods can utilize any DLL3 antibody, including for example, an anti-DLL3 antibody comprising three CDRs of a heavy chain variable region amino acid sequence of SEQ ID NO: 1-27 or 29.
An aspect of the invention is a humanized antibody which binds to human DLL3 protein comprising a light chain variable domain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% sequence identity to SEQ ID NO: 76-90 or 91.
The disclosed methods can utilize any DLL3 antibody, including for example, an anti-DLL3 antibody comprising three CDRs of a light chain variable region amino acid sequence of SEQ ID NO: 32-54 or 55.
An aspect of the invention is an antibody against MUC17. The antibody can be a fragment such as an antigen binding fragment (Fab) or a single chain variable fragment (Scfv).
An aspect of the invention is an agonist antibody that activates CD3, CD28 and/or CD137.
An aspect of the invention is a bispecific or trispecific molecule that includes an antibody (or fragment) against MUC17 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a method of treating an ailment such as cancer using an antibody (or fragment) against MUC17 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a method of treating an ailment that uses two or more of the bispecific or trispecific molecules described herein in combination with one another.
An aspect on the invention is a method of activating T-cell cytotoxicity against MUC17 expressing cells.
An aspect of the invention is a method of activating T-cell cytotoxicity using a bispecific or trispecific molecule the includes an antibody (or fragment) against MUC17 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a humanized antibody which binds to human MUC17 protein comprising a heavy chain variable domain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% sequence identity to SEQ ID NO: 127-245 or 246.
The disclosed methods can utilize any MUC17 antibody, including for example, an anti-MUC17 antibody comprising three CDRs of a heavy chain variable region amino acid sequence of SEQ ID NO: 92-111 or 112.
The disclosed methods can utilize any MUC17 antibody, including for example, an anti-MUC17 antibody comprising three CDRs of a light chain variable region amino acid sequence of SEQ ID NO: 113-125 or 126.
An aspect of the invention is a humanized antibody which binds to human MUC17 protein comprising a light chain variable domain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% sequence identity to SEQ ID NO: 147-168 or 169.
The disclosed methods can utilize any MUC17 antibody, including for example, an anti-MUC17 antibody comprising three CDRs of a heavy chain variable region amino acid sequence of SEQ ID NO: 92-111 or 112.
The disclosed methods can utilize any MUC17 antibody, including for example, an anti-MUC17 antibody comprising three CDRs of a light chain variable region amino acid sequence of SEQ ID NO: 113-125 or 126.
An aspect of the invention is a method of treating an ailment such as cancer using an antibody (or fragment) against CLDN18.2 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect on the invention is a method of activating T-cell cytotoxicity against CLDN18.2 expressing cells.
An aspect of the invention is a method of activating T-cell cytotoxicity using a bispecific or trispecific molecule the includes an antibody (or fragment) against CLDN18.2 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 or CD137.
An aspect of the invention is a method of activating T-cell cytotoxicity using a trispecific molecule the includes an antibody (or fragment) against CLDN18.2 paired with an antibody (or fragment) of an agonist antibody that activates CD3, CD28 and/or CD137.
The disclosed methods can utilize any CLDN18.2 antibody, including for example, an anti-CLDN18.2 antibody comprising three CDRs of a heavy chain variable region amino acid sequence of SEQ ID NO: 170-186 or 187.
An aspect of the invention is a humanized antibody which binds to human CLDN18.2 protein comprising a heavy chain variable domain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% sequence identity to SEQ ID NO: 197-205 or 206.
The disclosed methods can utilize any CLDN18.2 antibody, including for example, an anti-CLDN18.2 antibody comprising three CDRs of a light chain variable region amino acid sequence of SEQ ID NO: 188-195 or 196.
An aspect of the invention is a humanized antibody which binds to human CLDN18.2 protein comprising a light chain variable domain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% sequence identity to SEQ ID NO: 207-212 or 213.
The disclosed methods can utilize any CLDN18.2 antibody, including for example, an anti-CLDN18.2 antibody comprising three CDRs of a heavy chain variable region amino acid sequence of SEQ ID NO: 170-186 or 187.
The disclosed methods can utilize any CLDN18.2 antibody, including for example, an anti-CLDN18.2 antibody comprising three CDRs of a light chain variable region amino acid sequence of SEQ ID NO: 188-195 or 196.
Another aspect of the invention is a method of activating T-cell cytotoxicity using a trispecific molecule the includes more than one antibodies (or fragments) against DLL3, MUC17 and/or CLDN18.2 paired with one or more antibodies (or fragments) of an agonist antibodies that activate one or more of CD3, CD28 or CD137.
Another aspect of the invention is a trispecific molecule that includes (a) an anti-DLL3 antibody, an anti-Muc17 antibody or an anti-Cldn18.2 antibody, (b) a first antigen binding entity and (c) a second antigen binding entity. In one embodiment, the first antigen binding entity is comprised of an anti-CD28 scFv domain and the second antigen binding entity is comprised of an anti-CD3 Scfv-Fc domain. The first antigen binding entity and the second antigen binding entity form a heterodimer configuration. In one embodiment, the trispecific molecule includes a first chain of SEQ. ID NO: 347, a second chain of SEQ. ID NO: 276 and a third chain of SEQ. ID NO: 297. In another embodiment, the trispecific molecule includes a first chain of SEQ. ID NO: 430, a second chain of SEQ. ID NO: 276 and a third chain of SEQ. ID NO: 297. Alternatively, the trispecific molecule can include a first chain of SEQ. ID NO: 481, a second chain of SEQ. ID NO: 406 and a third chain of SEQ. ID NO: 413.
In some embodiments, the targeting domains are linked to one another by peptide bonds via peptide linkers or through covalent conjugates using appropriate crosslinking technologies known in the art.
In some embodiments, the targeting domains comprise antibody variable regions. In some embodiments, the targeting domains are in the form of a single domain antibody (sdAb), a fragment variable (Fv) heterodimer, a single chain Fv (scFv), a Fab fragment, a TriFab, or a combination thereof.
In some embodiments, the trispecific molecules are administered with a checkpoint inhibitor.
In some embodiments, the trispecific molecules are administered with an anti-PD1 and/or anti-POL1 antagonists.
Other features and advantages of aspects of the present invention will become apparent from the following more detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, the principles of aspects of the invention.



BRIEF DESCRIPTION OF THE DRAWINGS
The accompanying drawings illustrate aspects of the present invention. In such drawings:
FIG. 1A to 1R depict Scfv-scfv-Fc×Fab-Fc formats for the trispecific molecules of the invention.
FIG. 1A depicts an anti-CD3, anti-DLL3, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1B depicts an anti-CD3, anti-DLL3, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1C depicts an anti-CD28, anti-DLL3, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1D depicts an anti-CD28, anti-DLL3, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1E depicts an anti-CD137, anti-DLL3, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1F depicts an anti-CD137, anti-DLL3, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1G depicts an anti-DLL3, anti-CD3, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1H depicts an anti-DLL3, anti-CD3, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 11 depicts an anti-DLL3, anti-CD28, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1J depicts an anti-DLL3, anti-CD28, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1 K depicts an anti-DLL3, anti-CD137, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1 L depicts an anti-DLL3, anti-CD 137, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1 M depicts an anti-CD28, anti-CD3, anti-DLL3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1 N depicts an anti-CD137, anti-CD3, anti-DLL3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1O depicts an anti-CD 137, anti-CD28, anti-DLL3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1 P depicts an anti-CD3, anti-CD28, anti-DLL3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1Q depicts an anti-CD3, anti-CD137, anti-DLL3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 1 R depicts an anti-CD28, anti-CD137, anti-DLL3 (scfv-scfv-Fc×Fab-Fc)
FIG. 2A is a graph that demonstrates the binding of anti-DLL3 antibodies to human DLL3 expressing CHO cells.
FIG. 2B is a graph that demonstrates the binding of anti-DLL3 antibodies to CHO cells expressing cynomolgus DLL3.
FIG. 3 is a graph that demonstrates the binding of anti-DLL3 scfv-Fc to human DLL3 expressing CHO cells.
FIG. 4 is a graph that demonstrates CD28scfv-CD3scFv-Fc×DLL3-Fab-Fc trispecific molecule 328D3 activates PBMCs to secrete IFN gamma in the presence of CHO-DLL3 cells to higher levels than the CD3scfv-Fc×DLL3Fab-Fc bispecific molecule 3D1I or CD28scfv-Fc×DLL3Fab-Fc bispecific molecule 28D18.
FIG. 5 is a graph that demonstrates CD28scfv-CD3scFv-Fc×DLL3-Fab-Fc trispecific molecule 328D2 activates PBMCs to secrete IFNgamma in the presence of CHO-DLL3 cells to higher levels than the CD3scfv-Fc×DLL3Fab-Fc bispecific molecule 3D41 or CD28scfv-Fc×DLL3Fab-Fc bispecific molecule 28D15.
FIG. 6 is a graph that demonstrates CD28scfv-CD3scFv-Fc×DLL3-Fab-Fc trispecific molecule 328D1 activates PBMCs to secrete IFNgamma in the presence of CHO-DLL3 cells to higher levels than the CD3scfv-Fcx DLL3Fab-Fc bispecific molecule 3D221 or CD28scfv-Fcx DLL3Fab-Fc bispecific molecule 28D17
FIG. 7 is a graph that demonstrates CD28scfv-CD3scFv-Fcx DLL3-Fab-Fc trispecific molecules 328D4 and 328D5 activate PBMCs to secrete more IL-2 in the presence of NCI-H82 cells greater than the CD3scfv-Fcx DLL3Fab-Fc bispecific, 3D391.
FIG. 8A-8C depict bispecific and trispecific molecules with two CD137 Fab fragments.
FIG. 8A depicts a bispecific molecule of anti-CD137, anti-DLL3, anti-DLL3 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 8B depicts an anti-CD137, anti-DLL3, anti-CD3 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 8C depicts an anti-CD137, anti-DLL3, anti-CD3 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 9A-9F depict bispecific molecules and trispecific molecules with two CD137 scfv fragments.
FIG. 9A depicts an anti-DLL3, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fcx Fe-Scfv) molecule.
FIG. 9B depicts an anti-DLL3, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fcx Scfv-Fc-Scfv) molecule.
FIG. 9C depicts an anti-CD137, anti-CD137, anti-DLL3 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 9D depicts an anti-DLL3, anti-CD137, anti-CD137 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 9E depicts an anti-DLL3, anti-CD3, anti-CD137 (Fab-Fc-Scfv×Scfv-Fc-Scfv) molecule.
FIG. 9F depicts an anti-DLL3, anti-CD3, anti-CD137, anti-CD137 (Fab-Fc-Scfv-Scfv×Scfv-Fc) molecule.
FIG. 10A to 10R depict Scfv-scfv-Fc×Fab-Fc formats for the trispecific molecules of the invention.
FIG. 10A depicts an anti-CD3, anti-MUC17, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10B depicts an anti-CD3, anti-MUC17, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10C depicts an anti-CD28, anti-MUC17, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10D depicts an anti-CD28, anti-MUC17, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10E depicts an anti-CD137, anti-MUC17, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10F depicts an anti-CD137, anti-MUC17, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10G depicts an anti-MUC17, anti-CD3, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10H depicts an anti-MUC17, anti-CD3, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 10I depicts an anti-MUC17, anti-CD28, anti-CD137 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10J depicts an anti-MUC17, anti-CD28, anti-CD3 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10K depicts an anti-MUC17, anti-CD137, anti-CD3 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10L depicts an anti-MUC17, anti-CD137, anti-CD28 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10 M depicts an anti-CD28, anti-CD3, anti-MUC17 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10N depicts an anti-CD137, anti-CD3, anti-MUC17 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10O depicts an anti-CD137, anti-CD28, anti-MUC17 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10P depicts an anti-CD3, anti-CD28, anti-MUC17 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10Q depicts an anti-CD3, anti-CD137, anti-MUC17 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 10R depicts an anti-CD28, anti-CD137, anti-MUC17 (scfv-scfv-Fcx Fab-Fc) molecule.
FIG. 11A is a graph that demonstrates MUC17×CD3 Bispecifics bind CHOK1 cells expressing MUC17.
FIG. 11B is a graph that demonstrates MUC17×CD3 Bispecifics bind to ASPC1 tumor cells.
FIG. 12 graph showing hu1MU11A and hu1MU32A retain Muc17 binding activity in the Scfv format.
FIG. 13 is a graph that demonstrates CD3×CD28×Muc17 trispecific molecules have similar maximum killing of ASPC1 cells by PBMCs as the CD3 bispecific 3M62I.
FIG. 14 is a graph that demonstrates Muc17×CD3×cD28 Trispecific molecules 328M2,328M 3, 328M4, and 328M5 activate PBMCs in the presence of ASPC1 cells to secrete IL-2 at levels higher than that of the CD3 bispecific, 3M62I.
FIG. 15 is a graph that demonstrates CD3Scfv×CD28-scFv×Muc17Fab trispecific molecules 328M1 and 328M2 activate T-cells to kill MUC17-CHO cells better than the CD3 bispecific 3M55I, and similar to the combination of CD3 and CD28 bispecifics, 3M55I+28M3.
FIG. 16A-16B are graphs that demonstrate CD28scfv×CD3scfv×Muc17Fab trispecific 328M2 activates PBMCs to express CD25 only when in the presence of Muc17 expressing CHO cells and cannot activate without Muc17 target cells. In contrast the CD3scFv×CD28scFv×Muc17-Fab trispecific 328M1 is able to non-specifically activate cells in the absence of Muc17, similarly to the bispecific lacking Muc17 binding, 328-1.
FIG. 16A shows the expression of CD25(MFI) stimulated by increasing concentrations of the bispecific and trispecific molecules T cells when PBMCs are in the presence of MUCI 7 expressing CHO cells. FIG. 16B shows the expression of CD25(MFI) stimulated by increasing concentrations of the bispecific and trispecific molecules T cells when PBMCs are in the presence of parental CHO cells
FIGS. 17A and 17B are graphs that demonstrates CD137×CD3×Muc17 trispecific molecules 34M1 and 34M4 increase CD4 and CD8 T cell numbers when PBMCs are cultured with Muc17 expressing CHO cells to a greater level than the benchmark CD3 bispecific 3M8B7.
FIG. 18A-18D depict Muc17×CD137 and DLL3×CD137 bispecific molecules.
FIG. 19A-19C depict MUC17 bispecific and trispecific molecules with two CD137 Fab fragments.
FIG. 19A depicts an anti-CD137, anti-MUC17, anti-MUC17 (Fab-Fc-Scfv) molecule.
FIG. 19B depicts an anti-CD137, anti-MUC17, anti-CD3 (Fab-Fc-Scfv×Fab-Fc-Scfv) molecule.
FIG. 19C depicts an anti-CD137, anti-MUC17, anti-CD3 (Fab-Fc×Fc-Scfv-Scfv) molecule.
FIG. 19D-19I depict bispecific molecules and trispecific molecules with two anti-CD137 fragments.
FIG. 19D depicts an anti-CD137, anti-CD137, anti-MUC17, (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 19E depicts an anti-MUC17, anti-CD137, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 19F depicts an anti-MUC17, anti-CD137, anti-CD3, anti-CD137 (Fab-Fc-Scfv×Scfv-Fc-Scfv) molecule.
FIG. 19G depicts an anti-MUC17, anti-CD137, anti-CD137, anti-CD3 (Fab-Fc-Scfv-Scfv×Scfv-Fc) molecule.
FIG. 19H depicts an anti-MUC17, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc-Scfv×Fc-Scfv) molecule.
FIG. 19I depicts an anti-DLL3, anti-CD3, anti-CD137, anti-MUC17, anti-CD137 (scfv-scfv-Fc×Scfv-Fc-Scfv) molecule.
FIG. 20A to 20R depict Scfv-scfv-Fc×Fab-Fc formats for the trispecific molecules of the invention.
FIG. 20A depicts an anti-CD3, anti-CLDN18.2, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20B depicts an anti-CD3, anti-CLDN18.2, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20C depicts an anti-CD28, anti-CLDN18.2, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20D depicts an anti-CD28, anti-CLDN18.2, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20E depicts an anti-CD137, anti-CLDN18.2, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20F depicts an anti-CD137, anti-CLDN18.2, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20G depicts an anti-CLDN18.2, anti-CD3, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20H depicts an anti-CLDN18.2, anti-CD3, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20I depicts an anti-CLDN18.2, anti-CD28, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20J depicts an anti-CLDN18.2, anti-CD28, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20K depicts an anti-CLDN18.2, anti-CD137, anti-CD3 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20L depicts an anti-CLDN18.2, anti-CD137, anti-CD28 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20M depicts an anti-CD28, anti-CD3, anti-CLDN18.2 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20N depicts an anti-CD137, anti-CD3, anti-CLDN18.2 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20O depicts an anti-CD137, anti-CD28, anti-CLDN18.2 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20P depicts an anti-CD3, anti-CD28, anti-CLDN18.2 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20Q depicts an anti-CD3, anti-CD137, anti-CLDN18.2 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 20R depicts an anti-CD28, anti-CD137, anti-CLDN18.2 (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 21 is a graph that demonstrates trispecific molecules 328C1, 328C 2, 328C3, 328C4 activate T-cells to secrete IL-2 better to a greater level than the 3C27I CD3×CLDN18.2 bispecific alone.
FIG. 22 is a graph that demonstrates CD3×CD137×CLDN18.2 trispecific molecules 34C4 and 34C3 activate PBMCs in the presence of SNU-601 cells to express more CD25 than bispecific 3C18I.
FIG. 23 is a graph that demonstrates CD3×CD137×CLDN18.2 trispecific molecules 34C4 and 34C3 activate PBMCs in the presence of SNU-601 cells to secrete more IFNgamma than bispecific 3C18I alone.
FIG. 24 is a graph that demonstrates CD3×CD137×CLDN18.2 trispecific 34C3 increases CD8 T cell numbers beyond what is stimulated by the CD3 T cell engager 3C27I in the presence of CHO cells expressing CLDN18.2.
FIG. 25A-25C depict CLDN18.2 bispecific and trispecific molecules with two CD137 Fab or Scfv fragments.
FIG. 25A depicts an anti-CD137, anti-CLDN18.2, anti-CLDN (scfv-scfv-Fc×Fab-Fc) molecule.
FIG. 25B depicts an anti-CD137, anti-CLDN18.2, anti-CD3 trispecific molecule (Fab-Fc-Scfv×Fab-Fc-Scfv) molecule.
FIG. 25C depicts an anti-CD137, anti-CLDN18.2, anti-CD3 trispecific molecule (Fab-Fc-Scfv-Scfv×Fab-Fc)
FIG. 25D depicts an anti-CD137, anti-CD137, anti-CLDN18.2 bispecific molecule (Scfv-Scfv-Fc×Fab-Fc)
FIG. 25E depicts an anti-CLDN18.2, anti-CD137, anti-CD137, trispecific molecule (Scfv-Scfv-Fc×Fab-Fc)
FIG. 25F depicts an anti-CLDN18.2, anti-CD137, anti-CD3, anti-CD137 18.2 (Fab-Fc-Scfv×Scfv-Fc-Scfv) molecule.
FIG. 25G depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (Fab-Fc-Scfv-Scfv×Scfv-Fc) molecule.
FIG. 25H depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc-Scfv×Fc-Scfv) molecule.
FIG. 25I depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc×Scfv-Fc-Scfv) molecule.



DEFINITIONS
Reference in this specification to “one embodiment/aspect” or “an embodiment/aspect” means that a particular feature, structure, or characteristic described in connection with the embodiment/aspect is included in at least one embodiment/aspect of the disclosure. The use of the phrase “in one embodiment/aspect” or “in another embodiment/aspect” in various places in the specification are not necessarily all referring to the same embodiment/aspect, nor are separate or alternative embodiments/aspects mutually exclusive of other embodiments/aspects. Moreover, various features are described which may be exhibited by some embodiments/aspects and not by others. Similarly, various requirements are described which may be requirements for some embodiments/aspects but not other embodiments/aspects. Embodiment and aspect can in certain instances be used interchangeably.
The terms used in this specification generally have their ordinary meanings in the art, within the context of the disclosure, and in the specific context where each term is used. Certain terms that are used to describe the disclosure are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the disclosure. It will be appreciated that the same thing can be said in more than one way.
Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein. Nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only, and is not intended to further limit the scope and meaning of the disclosure or of any exemplified term. Likewise, the disclosure is not limited to various embodiments given in this specification.
Without intent to further limit the scope of the disclosure, examples of instruments, apparatus, methods and their related results according to the embodiments of the present disclosure are given below. Note that titles or subtitles may be used in the examples for convenience of a reader, which in no way should limit the scope of the disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In the case of conflict, the present document, including definitions, will control.
As applicable, the terms “about” or “generally”, as used herein in the specification and appended claims, and unless otherwise indicated, means a margin of +/−20%. Also, as applicable, the term “substantially” as used herein in the specification and appended claims, unless otherwise indicated, means a margin of +/−10%. It is to be appreciated that not all uses of the above terms are quantifiable such that the referenced ranges can be applied.
The term “subject” or “patient” refers to any single animal, more preferably a mammal (including such non-human animals as, for example, dogs, cats, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human primates) for which treatment is desired. Most preferably, the patient herein is a human. In an embodiment, a “subject” of diagnosis or treatment is a prokaryotic or a eukaryotic cell, a tissue culture, a tissue or an animal, e.g. a mammal, including a human.
As used herein, the term “comprising” is intended to mean that the compositions and methods include the listed elements, but do not exclude other unlisted elements. “Consisting essentially of” when used to define compositions and methods, excludes other elements that alters the basic nature of the composition and/or method, but does not exclude other unlisted elements. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace amounts of elements, such as contaminants from any isolation and purification methods or pharmaceutically acceptable carriers, such as phosphate buffered saline, preservatives, and the like, but would exclude additional unspecified amino acids. “Consisting of” excludes more than trace elements of other ingredients and substantial method steps for administering the compositions described herein. Embodiments defined by each of these transition terms are within the scope of this disclosure and the inventions embodied therein.
The term “tri-specific monoclonal antibody” or “TSMAb” refers to an antibody that can simultaneously engage three different types of epitopes.
The term “active agent” or “active ingredient” refers to a substance, compound, or molecule, which is biologically active or otherwise, induces a biological or physiological effect on a subject to which it is administered to. In other words, “active agent” or “active ingredient” refers to a component or components of a composition to which the whole or part of the effect of the composition is attributed. An active agent can be a primary active agent, or in other words, the component(s) of a composition to which the whole or part of the effect of the composition is attributed. An active agent can be a secondary agent, or in other words, the component(s) of a composition to which an additional part and/or other effect of the composition is attributed.
In an embodiment, a “pharmaceutical composition” is intended to include the combination of an active agent, such as an anti-DLL3, anti-MUC17 or anti-CLDN18.2 antibody and antibody conjugates, with a carrier, inert or active, in a sterile composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo. In one aspect, the pharmaceutical composition is substantially free of endotoxins or is non-toxic to recipients at the dosage or concentration employed.
In an embodiment, “an effective amount” refers, without limitation, to the amount of the defined component sufficient to achieve the desired therapeutic result. In an embodiment, that result can be effective cancer treatment.
In an embodiment, as used herein, the terms “treating,” “treatment” and the like are used herein, without limitation, to mean obtaining a desired pharmacologic and/or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disorder or sign or symptom thereof, and/or may be therapeutic in terms of amelioration of the symptoms of the disease or infection, or a partial or complete cure for a disorder and/or adverse effect attributable to the disorder.
As used herein, the term “recombinant” refers to polypeptides or polynucleotides that do not exist naturally and which may be created by combining polynucleotides or polypeptides in arrangements that would not normally occur together. The term can refer to a polypeptide produced through a biological host, selected from a mammalian expression system, an insect cell expression system, a yeast expression system, and a bacterial expression system.
The term “Delta-like 3” or “DLL3” refers to a protein which in humans is encoded by the DLL3 gene. Mutations in the gene cause the autosomal recessive genetic disorder Jarcho-Levin syndrome. DLL3 is expressed normally on the inside of cells and at low levels on normal tissues. However, lung tumor cells overexpress the gene and cell surface DLL3 levels are increased.
The term “Mucin 17” or “MUC17” refers to a member of the mucin family that includes more than 20 members. Mucins are large, highly glycosylated membrane bound proteins. They are expressed almost exclusively in the intestine. Their general function is to protect epithelial cells from their environment, as well as to regulate proliferation and survival of cells. MUC17 is highly expressed in pancreatic adenocarcinoma tissue (at protein level). MUC17 is expressed in pancreatic, appendiceal, and some colon cancers. Its expression is not detectable in normal pancreas, in pancreatitis or in cell lines derived from other cancers.
The term “Claudin-18” or “CLD 18” refers to a protein that in humans is encoded by the CLDN18 gene. CLDN18 belongs to the large claudin family of proteins, which form tight junction strands in epithelial cells. “Claudin 18.2” or “CLDN18.2” denotes isoform 2 which is abundant in tumors, particularly those of the gastric system.
The term “CD” or “cluster of differentiation molecules” refers to cell surface markers that are useful for the identification and characterization of leukocytes such as CD3, CD28 and CD137. CD3 is the signaling component of the T cell receptor (TCR) complex. Because CD3 is required for T cell activation, drugs (often monoclonal antibodies) that target it are being investigated as immunostimulants for the treatment of cancer.
CD28 is the major costimulatory molecule required in the generation of T cell-mediated immune responses. Upon interaction with its ligands CD80 and CD86, CD28 transduces activation signals that lead to the expression of anti-apoptotic proteins and enhance the synthesis of several cytokines including IL-2. CD28 costimulatory receptor is present on all T-cells. Agonist antibodies directed against CD28 have led to severe adverse events in the clinic, in contrast to antibodies directed against the other CD28 family members CTLA-4, PD-1, or their B7 ligands, which function as checkpoint inhibitors to overcome tumor immune tolerance and can be used in cancer immunotherapy.
CD137 is a member of the tumor necrosis factor (TNF) receptor family. Its alternative names are tumor necrosis factor receptor superfamily member 9 (TNFRSF9), 4-1BB and induced lymphocyte activation. Agonistic anti-CD137 antibody acts as an activating costimulatory molecule especially important for effector/memory T cells and promotes the survival and proliferation of T lymphocytes. For example, BBK-4, Urelumab and Utomilumab (PF-05082566) targets this receptor to stimulate a more intense immune system attack on cancers.
PD-1 (Programmed cell death protein 1 or CD279) is a protein on the surface of cells that has a role in regulating the immune system's response to the cells of the human body by down-regulating the immune system and promoting self-tolerance by suppressing T cell inflammatory activity. Engagement of PD-1 by either of its ligands, PD-L1 or PD-L2, on an adjacent cell inhibits TCR signaling and TCR-mediated proliferation, transcriptional activation and cytokine production. This prevents autoimmune diseases, but it can also prevent the immune system from killing cancer cells. Therapeutic antibodies designed to block the PD-1/PD-L1 interaction have potential for the treatment of cancer.
Programmed death-ligand 1 (PD-L1) also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1) is a protein that in humans is encoded by the CD274 gene. PD-L1 binds to its receptor, PD-1, found on activated T cells, B cells, and myeloid cells, to modulate activation or inhibition.
As used herein, the term “antibody” refers to a polypeptide or a polypeptide complex that specifically recognizes and binds to an antigen through one or more immunoglobulin variable regions. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. Typically, the antigen-binding region of an antibody will be most critical in specificity and affinity of binding and is encoded by the variable domain. An antibody can be a whole antibody, an antigen binding fragment or a single chain thereof.
An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one “light” (about 25 kD) and one “heavy” chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to variable domains of the light and heavy chain respectively.
Antibodies exist, e.g., as intact immunoglobulins or as a number of well-characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)′2, a dimer of Fab which itself is a light chain VL-CL joined to VH-CH1 by a disulfide bond. While various antibody fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that such fragments may be synthesized de novo either chemically or by using recombinant DNA methodology. Thus, the term antibody, as used herein, also includes antibody fragments either produced by the modification of whole antibodies, or those synthesized de novo using recombinant DNA methodologies (e.g., single chain Fv) or those identified using phage display libraries (see, e.g., McCafferty et al., Nature 348:552-554 (1990)).
Accordingly, in either aspect of the invention, the term antibody also embraces minibodies, scFvs, diabodies, triabodies and the like. ScFvs and Diabodies are small bivalent biospecific antibody fragments with high avidity and specificity. Their high signal to noise ratio is typically better due to a better specificity and fast blood clearance increasing their potential for diagnostic and therapeutic targeting of specific antigen (Sundaresan et al., J Nucl Med 44:1962-9 (2003). In addition, these antibodies are advantageous because they can be engineered if necessary as different types of antibody fragments ranging from a small single chain Fv (scFv) to an intact IgG with varying isoforms (Wu & Senter, Nat. Biotechnol. 23:1137-1146 (2005)). In some embodiments, the antibody fragment is part of a scFv-scFv or diabody. In some embodiments, in either aspect, the invention provides high avidity antibodies for use according to the invention.
The term “agonist antibody” refers to an antibody that stimulates or activates an organ. An antibody can act as an agonist of a receptor, essentially replacing the activity of the normal ligand. The agonist activity can occur when the antibody binds the receptor in a manner that mimics the binding of the physiological ligand resulting in antibody-mediated agonism. For example, agonistic antibodies against the thyrotropin receptor in Grave's disease stimulate the thyroid gland to release thyroid hormones that produce hyperthyroidism. Agonistic antibodies may also stimulate when clustered, either via the Fe portion of the antibody engaging an Fc receptor in trans or cis, or through antigen mediated clustering. The latter clustering mechanism requires antigen engagement by one half of a bispecific or trispecific molecule and engagement of the stimulatory receptor by the second half of a bispecific or trispecific molecule. Exemplary stimulatory receptors are CD3, CD28 and 4-1BB, which stimulate T cells.
The terms “antibody fragment” or “antigen-binding fragment” are used with reference to a portion of an antibody, such as Fab′, Fab, Fv, scFv and the like. Regardless of structure, an antibody fragment binds with the same antigen that is recognized by the intact antibody. The term “antibody fragment” also includes diabodies and any synthetic or genetically engineered proteins comprising immunoglobulin variable regions that act like an antibody by binding to a specific antigen to form a complex.
The term “antigen-binding fragment” or “Fab” refers to a region on an antibody that binds to antigens. It includes one constant and one variable domain of each of the heavy and the light chain (i.e. four domains: VH, CH1, VL and CL1). The variable domain contains the paratope (the antigen-binding site), that includes a set of complementary determining regions at the amino terminal end of the monomer. Each arm of the Y thus binds an epitope on the antigen.
The term “Fc region” or “fragment crystallizable region” refers to the tail region of an antibody CH2-CH3 that interacts with cell surface receptors called Fc receptors and some proteins of the complement system. This “effector function” allows antibodies to activate the immune system leading to cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and/or complement dependent cytotoxicity (CDC). ADCC and ADCP are mediated through the binding of the Fc to Fc receptors on the surface of cells of the immune system. CDC is mediated through the binding of the Fe with proteins of the complement system, (e.g. C1q).
In IgG, IgA and IgD antibody isotypes, the Fe region has two identical protein fragments, derived from the second and third constant domains of the antibody's two heavy chains. IgM and IgE Fc regions have three heavy chain constant domains (CH domains 2-4) in each polypeptide chain whereas IgG is composed of 2 CH domains, 2 and 3. The Fc regions of IgGs bear a highly conserved N-glycosylation site. Glycosylation of the Fc fragment is essential for Fc receptor-mediated activity. The N-glycans attached to this site are predominantly core-fucosylated diantennary structures of the complex type. In addition, small amounts of these N-glycans also bear bisecting GlcNAc and a-2,6 linked sialic acid residues.
A particular IgG subclass can be preferred for a particular use. For example, IgG1 is more effective than IgG2 and IgG4 at mediating ADCC and CDC. Thus, IgG2 Fc can be preferred when effector function is undesirable. However, IgG2 Fe-containing molecules are generally more difficult to manufacture and can be less stable than IgG1 Fc-containing molecules. Further, the effector function of an antibody can be increased, or decreased, by introducing one or more mutations into the Fc (see, for example, Strohl, Curr. Opin. Biotech., 20:685-691, 2009). Exemplary IgG1 Fc molecules having increased effector function include those having the following substitutions:

    
    
        S239D/I332E, S239D/A330St1332E, S2390/A330L/1332E, S298A/0333A/K334A, P2471/A339D, P2471/A339Q, 0280H/K290S, 0280H/K290S/S298D, 0280H/K290S/S298V, F243L/R292P/Y300L, F243L/R292P/Y300L/P396L, F243L/R292P/Y300L/V3051/P396L, G236A/S239D/I332E, K326A/E333A, K326W/E333S, K290E/S298G/T299A, K290N/S298G/T299A, K290E/S298G/T299A/K326E, K290N/S298G/T299A/K326E
    
    


Fucosylation is another method of increasing effector function of IgG Fe-containing proteins. Removal of the core fucose from the biantennary complex-type oligosaccharides attached to the Fc greatly increases ADCC effector function without altering antigen binding or CDC effector function. There are different ways to reduce or abolish fucosylation of Fe-containing molecules. These include recombinant expression in certain mammalian cell lines including a FUT8 knockout cell line, variant CHO line Lec13, rat hybridoma cell line YB2/0, a cell line comprising a small interfering RNA specifically against the FUT8 gene, and a cell line co-expressing β-1,4-N-acetylglucosaminyltransferase Ill and Golgi a-mannosidase II. Alternatively, the Fc-containing molecule can be expressed in a non-mammalian cell such as a plant cell, yeast, or prokaryotic cell, e.g., E. coli. 
It may be desirable to decrease effector function. Exemplary Fc molecules having decreased effector function include those having the following substitutions:

    
    
        N297A or N297Q (IgG1), L234A/L235A (IgG1), V234A/G237A (IgG2), L235A/G237A/E318A (IgG4), H268Q/V309L/A330S/A331S (IgG2), C220S/C226S/C229S/P238S (IgG1), C226S/C229S/E233P/L234V/L235A (IgG1), L234F/L235E/P331S (IgG1), S267E/L328F (IgG1)
    
    


Both the light and heavy chains are divided into regions of structural and functional homology. The terms “constant” and “variable” are used functionally. In this regard, it will be appreciated that the variable domains of both the light (VL) and heavy (VH) chain portions determine antigen recognition and specificity. Conversely, the constant domains of the light chain (CL) and the heavy chain (CHI, CH2 and CH3) confer important biological properties such as secretion, transplacental mobility, Fe receptor binding, complement binding, and the like. By convention, the numbering of the constant region domains in conventional antibodies increases as they become more distal from the antigen-binding site or amino-terminus of the antibody. In conventional antibodies, the N-terminal portion is a variable region and at the C-terminal portion is a constant region; the CH3 and CL domains comprise the carboxyterminus of the heavy and light chain, respectively.
As used herein, the term “heavy chain constant region” includes amino acid sequences derived from an immunoglobulin heavy chain. A polypeptide comprising a heavy chain constant region comprises at least one of: a CHI domain, a hinge (e.g., upper, middle, and/or lower hinge region) domain, a CH2 domain, a CH3 domain, or a variant or fragment thereof. For example, an antigen-binding polypeptide for use in the disclosure may comprise a polypeptide chain comprising a CHI domain; a polypeptide chain comprising a CHI domain, at least a portion of a hinge domain, and a CH2 domain; a polypeptide chain comprising a CHI domain and a CH3 domain; a polypeptide chain comprising a CHI domain, at least a portion of a hinge domain, and a CH3 domain, or a polypeptide chain comprising a CHI domain, at least a portion of a hinge domain, a CH2 domain, and a CH3 domain. In some embodiments, a polypeptide of the disclosure comprises a polypeptide chain comprising a CH3 domain. Further, an antibody for use in the disclosure may lack at least a portion of a CH2 domain (e.g., all or part of a CH2 domain). It should be understood that the heavy chain constant region may be modified such that they vary in amino acid sequence from the naturally occurring immunoglobulin molecule.
The heavy chain constant region of an antibody disclosed herein may be derived from different immunoglobulin molecules. For example, a heavy chain constant region of a polypeptide may comprise a CHI domain derived from an IgG1 molecule and a hinge region derived from an IgG3 molecule. In another example, a heavy chain constant region can comprise a hinge region derived, in part, from an IgG1 molecule and, in part, from an IgG3 molecule. In another example, a heavy chain portion can comprise a chimeric hinge derived, in part, from an IgG1 molecule and, in part, from an IgG4 molecule.
As used herein, the term “light chain constant region” includes amino acid sequences derived from antibody light chain. Preferably, the light chain constant region comprises at least one of a constant kappa domain or constant lambda domain. A “light chain heavy chain pair” refers to the collection of a light chain and heavy chain that can form a dimer through a disulfide bond between the CL domain of the light chain and the CHI domain of the heavy chain.
The subunit structures and three-dimensional configurations of the constant regions of the various immunoglobulin classes are well known. As used herein, the term “VH domain” includes the amino terminal variable domain of an immunoglobulin heavy chain and the term “CHI domain” includes the first (most amino terminal) constant region domain of an immunoglobulin heavy chain. The CHI domain is adjacent to the VH domain and is amino terminal to the hinge region of an immunoglobulin heavy chain molecule.
As used herein the term “CH2 domain” includes the portion of a heavy chain molecule that extends, e.g., from about residue 244 to residue 360 of an antibody using conventional numbering schemes (residues 244 to 360, Kabat numbering system; and residues 231-340, EU numbering system). The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. The CH3 domain extends from the CH2 domain to the C-terminal of the IgG molecule and comprises approximately 108 residues.
As used herein, the term “hinge region” includes the portion of a heavy chain molecule that joins the CHI domain to the CH2 domain. This hinge region comprises approximately 25 residues and is flexible, thus allowing the two N-terminal antigen-binding regions to move independently. Hinge regions can be subdivided into three distinct domains: upper, middle, and lower hinge domains.
The term “bi-specific monoclonal antibody” or “BSMAb” refers to an antibody that can simultaneously engage two different types of epitopes on the same target or on different targets. An advantage is their ability to redirect specific polyclonal immune cells (e.g. T cells and NK cells) to tumor cells to enhance tumor killing. These antibodies can be divided into two types: IgG like bispecific antibodies which carry an Fc region and therefore retain Fe-mediated effector functions and the non-IgG like formats which rely on their antigen binding capacity to exert their effects. Recombinant techniques have also led to the creation of small fragment molecules. Single chain variable fragments from two different monoclonal antibodies can be combined to form bivalent bispecific antibodies. Examples include bispecific T cell engagers (BiTEs), tandem single chain variable fragments (taFvs), diabodies (Dbs), single chain diabodies (scDbs), and triple bodies. These scFv based antibody fragments have high tumor specificity and tumor penetration due to their small size (ranging from 50 to 60 kDa).
The term “tri-specific monoclonal antibody” or “TSMAb” refers to an antibody that can simultaneously engage three different types of epitopes on the same target or on different targets.
The term “scFv” or “scFv fragment antibody” refers to a small molecular antibody, consisting of VH and VL domains, either in the configuration of VL-VH or VH-VL, with a linker region between them. The scFv fragment antibody can more easily penetrate blood vessel wall and the solid tumor, which makes it a preferred carrier of targeting drugs.
The term “scFvs” or “single-chain variable fragment” refers to divalent (or bivalent) single-chain variable fragments (di-scFvs, bi-scFvs) that can be engineered by linking two scFvs. This can be done by producing a single peptide chain with two VH and two VL regions, yielding tandem scFvs, also known as scFv-scFv molecules. Another possibility is the creation of scFvs with linker peptides that are too short for the two variable regions to fold together (about five amino acids), forcing scFvs to dimerize. This type is known as diabodies.
The term “humanized antibody” refers to an antibody from non-human species whose protein sequences have been modified to increase its similarity to antibody variants produced naturally in humans. The process of “humanization” is usually applied to monoclonal antibodies developed for administration to humans (e.g. antibodies developed as anti-cancer drugs). Humanization can be necessary when the process of developing a specific antibody involves generation in a non-human immune system (such as that in mice).
Bispecific or trispecific antibodies can be generated by chemical cross-linking or by the hybrid hybridoma technology. Alternatively, bispecific or trispecific antibody molecules can be produced by recombinant techniques, for example by linking 2 scFv molecules together with a short linker. For example, VH1-Linker1-VL1-Linker2-VH2-Linker3-VL2. With Linker1 and Linker3 having lengths between 15-30 amino acids and Linker2 being 5-10 amino acids in length. Linkers may be composed of a variety of amino acids, for example repeating units of GGGGS (SEQ ID NO: 504, GKPGS (SEQ ID NO: 505, GEPGS (SEQ ID NO: 506, and/or GGPGS (SEQ ID NO: 507. Dimerization across 2 scFv molecules can be promoted by reducing the length of the linker joining the VH and the VL domain from about 15 amino acids, routinely used to produce scFv fragments, to about 5 amino acids. These linkers favor intrachain assembly of the VH and VL domains, with the configuration VH1-linker1-VL2-Linker2-VH2-Linker 3VL1 and linkers 1 and 3 being 5 amino acids in length. Any suitable short linker can be used. Thus, two fragments assemble into a dimeric molecule. Further reduction of the linker length to 0-2 amino acids can generate trimeric (triabodies) or tetrameric (tetrabodies) molecules.
A “chimeric antibody” is an antibody molecule in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species, or an entirely different molecule which confers new properties to the chimeric antibody, e.g., an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity.
One of the challenges for efficiently producing bispecific antibody preparations concerns reducing the formation of homodimeric molecules in favor of heterodimeric molecules, when co-expressing chains of different binding specificities. A “heterodimeric antibody” can utilize the “knobs-into-holes” or “charge-pair” formats to preferentially promote correct association of the 2 molecules to form a heterodimer with 2 specificities. These formats are specific to the heavy chain Fc part of the constant region in antibodies. For the knob-into-holes format, the “knobs” part is engineered by replacing a small amino acid with a larger one. It fits into the “hole,” which is engineered by replacing a large amino acid with a smaller one. Introduction of T366W mutations in the first Fc creates the “knob” and introduction of T366S, L368A, and Y407V mutations in the second Fc creates the “hole” (numbering of the residues according to the Kabat EU numbering system). For the charge pair format, heterodimerization is favored through stabilizing ionic interactions by introducing interfacing charge residues in the opposing Fc domains. For example, D356K, E357K, and D399K in a first Fc domain, and the mutations K370E, K409D, and K439E into a second Fc domain, or combination thereof. For example, K392D and K409D mutations in a first Fc chain, and D399K and D356K mutations in a second Fc chain, K409E in the first Fc and D399K in the Fc, K409E in the first Fc and D399R in the second Fc, K409D in the first Fc and D0399K in the second Fc, K409D in the first Fc and D399R in the second Fc, K392E in the first Fe and D399R in the second Fe, K392E in the first Fe and 0399K in the second Fe, K392D in the first Fc and D399R in the second Fc, K392D in the first Fc and 0399K in the second Fc, K409D and K360D in the first Fc and D399K and D356K in the second Fc, K409D and K370D in the first Fc and D399K and E357K in the second Fc, K409D and K392D in the first Fc and D399K, D356K, and E357K in the second Fc, K409D and K392D in the first Fc and D399K in the second Fc, K409D and K392D in the first Fc and D399K and D356K in the second Fc, K409D and K392D in the first Fc and D399K and E357K in the second Fc, K409D and K370D in the first Fc and D399K and D357K in the second Fc, D399K in the first Fc and K409D and K360D in the second Fc, and/or K409D and K439D in the first Fc and D399K and D356K in the second Fc, numbered according to the Kabat EU numbering system. Additionally, cysteines may be introduced to stabilize the pairing of heterodimers, for example S234C in the first Fc and Y349C in the second Fc or Y349C in the first Fc and S344C in the second Fc.
The phrase “specifically (or selectively) binds” to an antibody or “specifically (or selectively) immunoreactive with,” when referring to a protein or peptide, refers to a binding reaction that is determinative of the presence of the protein, often in a heterogeneous population of proteins and other biologics. Thus, under designated immunoassay conditions, the specified antibodies bind to a particular protein at least two times the background and more typically more than 10 to 100 times background. Specific binding to an antibody under such conditions requires an antibody that is selected for its specificity for a particular protein. For example, polyclonal antibodies can be selected to obtain only those polyclonal antibodies that are specifically immunoreactive with the selected antigen and not with other proteins. This selection may be achieved by subtracting out antibodies that cross-react with other molecules. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein (see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual(1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity).
An “immune response” refers to the action of a cell of the immune system (for example, T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells and neutrophils) and soluble macromolecules produced by any of these cells or the liver (including Abs, cytokines, and complement) that results in selective targeting, binding to, damage to, destruction of, and/or elimination from a vertebrate's body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in cases of autoimmunity, neurodegeneration or pathological inflammation, normal human cells or tissues.
An “immunoregulator” refers to a substance, an agent, a signaling pathway or a component thereof that regulates an immune response. “Regulating,” “modifying” or “modulating” an immune response refers to any alteration in a cell of the immune system or in the activity of such cell. Such regulation includes stimulation or suppression of the immune system which may be manifested by an increase or decrease in the number of various cell types, an increase or decrease in the activity of these cells, or any other changes which can occur within the immune system. Both inhibitory and stimulatory immunoregulators have been identified, some of which may have enhanced function in the cancer, infectious disease or neurodegenerative microenvironment.
A cytotoxic T cell (also known as TC, cytotoxic T lymphocyte, CTL, T-killer cell, cytolytic T cell, CD8+ T-cell or killer T cell) is a T lymphocyte (a type of white blood cell) that kills cancer cells, cells that are infected (particularly with viruses), or cells that are damaged in other ways.
The term “immunotherapy” refers to the treatment of a subject afflicted with, or at risk of contracting or suffering a recurrence of, a disease by a method comprising inducing, enhancing, suppressing or otherwise modifying an immune response. “Treatment” or “therapy” of a subject refers to any type of intervention or process performed on, or the administration of an active agent to, the subject with the objective of reversing, alleviating, ameliorating, inhibiting, slowing down or preventing the onset, progression, development, severity or recurrence of a symptom, complication, condition or biochemical indicia associated with a disease.
“Potentiating an endogenous immune response” means increasing the effectiveness or potency of an existing immune response in a subject. This increase in effectiveness and potency may be achieved, for example, by overcoming mechanisms that suppress the endogenous host immune response or by stimulating mechanisms that enhance the endogenous host immune response.
Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a pre-sequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a pre-protein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, “operably linked” means that the DNA sequences being linked are near each other, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.
“Conservatively modified variants” applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, conservatively modified variants refers to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are “silent variations,” which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence with respect to the expression product, but not with respect to actual probe sequences.
The terms “identical” or percent “identity,” in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection. Such sequences are then said to be “substantially identical.” This definition also refers to, or may be applied to, the compliment of a test sequence. The definition also includes sequences that have deletions and/or additions, as well as those that have substitutions. As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 amino acids or nucleotides in length.
For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Preferably, default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
A “comparison window,” as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to the full length of the reference sequence, usually about 25 to 100, or 50 to about 150, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 supplement)).
A preferred example of algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990), respectively. BLAST and BLAST 2.0 are used, with the parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=−4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=−4, and a comparison of both strands.
“Nucleic acid” refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, and complements thereof. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)). The term nucleic acid is used interchangeably with gene, cDNA, mRNA, oligonucleotide, and polynucleotide.
A particular nucleic acid sequence also implicitly encompasses “splice variants.” Similarly, a particular protein encoded by a nucleic acid implicitly encompasses any protein encoded by a splice variant of that nucleic acid. “Splice variants,” as the name suggests, are products of alternative splicing of a gene. After transcription, an initial nucleic acid transcript may be spliced such that different (alternate) nucleic acid splice products encode different polypeptides. Mechanisms for the production of splice variants vary, but include alternate splicing of exons. Alternate polypeptides derived from the same nucleic acid by read-through transcription are also encompassed by this definition. Any products of a splicing reaction, including recombinant forms of the splice products, are included in this definition. An example of potassium channel splice variants is discussed in Leicher et al., J. Biol. Chem. 273(52):35095-35101 (1998).
The term “heterologous” when used with reference to portions of a nucleic acid indicates that the nucleic acid comprises two or more subsequences that are not found in the same relationship to each other in nature. For instance, the nucleic acid is typically recombinantly produced, having two or more sequences from unrelated genes arranged to make a new functional nucleic acid, e.g., a promoter from one source and a coding region from another source. Similarly, a heterologous protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (e.g., a fusion protein).
As used herein, the term “prevention” means all of the actions by which the occurrence of the disease is restrained or retarded.
As used herein, the term “treatment” means all of the actions by which the symptoms of the disease have been alleviated, improved or ameliorated. In the present specification, “treatment” means that the symptoms of cancer, neurodegeneration, or infectious disease are alleviated, improved or ameliorated by administration of the antibodies disclosed herein.
The term “administration” refers to the introduction of an amount of a predetermined substance into a patient by a certain suitable method. The composition disclosed herein may be administered via any of the common routes, as long as it is able to reach a desired tissue, for example, but is not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrapulmonary, or intrarectal administration. However, since peptides are digested upon oral administration, active ingredients of a composition for oral administration should be coated or formulated for protection against degradation in the stomach.
The term “subject” refers to those suspected of having or diagnosed with cancer, a neurodegenerative or an infectious disease. However, any subject to be treated with the pharmaceutical composition disclosed herein is included without limitation. The pharmaceutical composition including an anti-DLL3 antibody disclosed herein is administered to a subject suspected of having cancer, a neurodegenerative or an infectious disease.
Construction of suitable vectors containing the desired sequences and control sequences employs standard ligation and restriction techniques, which are well understood in the art (see Maniatis et al., in Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York (1982)). Isolated plasmids, DNA sequences, or synthesized oligonucleotides are cleaved, tailored, and re-ligated in the form desired.
For preparation of antibodies, e.g., recombinant, monoclonal, or polyclonal antibodies, many techniques known in the art can be used (see, e.g., Kohler & Milstein, Nature 256:495-497 (1975); Kozbor et al., Immunology Today 4:72 (1983); Cole et al., in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96 (1985); Coligan, Current Protocols in Immunology (1991); Harlow & Lane, Antibodies, A Laboratory Manual (1988); and Goding, Monoclonal Antibodies: Principles and Practice (2d ed. 1986)). The genes encoding the heavy and light chains of an antibody of interest can be cloned from a cell, e.g., the genes encoding a monoclonal antibody can be cloned from a hybridoma and used to produce a recombinant monoclonal antibody. Gene libraries encoding heavy and light chains of monoclonal antibodies can also be made from hybridoma or plasma cells. Random combinations of the heavy and light chain gene products generate a large pool of antibodies with different antigenic specificity (see, e.g., Kuby, Immunology (3rd ed. 1997)). Techniques for the production of single chain antibodies or recombinant antibodies (U.S. Pat. Nos. 4,946,778, 4,816,567) can be adapted to produce antibodies to polypeptides of this invention. Also, transgenic mice, or other organisms such as other mammals, may be used to express humanized or human antibodies (see, e.g., U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, Marks et al., Bio/Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859 (1994); Morrison, Nature 368:812-13 (1994); Fishwild et al., Nature Biotechnology 14:845-51 (1996); Neuberger, Nature Biotechnology 14:826 (1996); and Lonberg & Huszar, Intern. Rev. Immunol. 13:65-93 (1995)). Alternatively, phage display technology can be used to identify antibodies and heteromeric Fab fragments that specifically bind to selected antigens (see, e.g., McCafferty et al., Nature 348:552-554 (1990); Marks et al., Biotechnology 10:779-783 (1992)). Antibodies can also be made bispecific, i.e., able to recognize two different antigens (see, e.g., WO 93/08829, Traunecker et al., EMBO J. 10:3655-3659 (1991); and Suresh et al., Methods in Enzymology 121-210 (1986)). Antibodies can also be heteroconjugates, e.g., two covalently joined antibodies, or immunotoxins (see, e.g., U.S. Pat. No. 4,676,980, WO 91/00360; and WO 92/200373).
Methods for humanizing antibodies are well known in the art. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can be essentially performed following the method of Winter and co-workers (see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988) and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
The term “cancer” refers to human cancers and carcinomas, sarcomas, adenocarcinomas, etc., including solid tumors, kidney, breast, lung, kidney, bladder, urinary tract, urethra, penis, vulva, vagina, cervical, colon, ovarian, prostate, pancreas, stomach, brain, head and neck, skin, uterine, testicular, esophagus, and liver cancer. Additional cancers include, for example, Hodgkin's Disease, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, small-cell lung tumors, primary brain tumors, stomach cancer, colon cancer, malignant pancreatic insulanoma, malignant carcinoid, premalignant skin lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, cervical cancer, endometrial cancer, and adrenal cortical cancer.
The term “checkpoint inhibitor” or “immune checkpoint inhibitor” refers to an agent such as a drug that inhibits/blocks the inhibitory checkpoint molecules. Some cancers can protect themselves from attack by stimulating immune checkpoint targets. Checkpoint therapy can block inhibitory checkpoints, restoring immune system function.
The term “immune checkpoint regulator” refers to receptors and their associated ligands, which together provide a means for inhibiting or stimulating signaling pathways that otherwise lead to T-cell activation. Immune checkpoint regulators include TIGIT and its CD155 ligand, PVR; PD-1 and its ligands, PD-L1 and PD-L2; CTLA-4 and its ligands, B7-1 and B7-2; TIM-3 and its ligand, Galectin-9; LAG-3 and its ligands, including liver sinusoidal endothelial cell lectin (LSECtin) and Galectin-3; CD122 and its CD122R ligands; CD70, B7H3, B and T lymphocyte attenuator (BTLA), and VISTA.
The term “checkpoint regulator antagonist,” “immune checkpoint binding antagonist” and “immune checkpoint antagonist” refer to a class of agents that interfere with (or inhibit) the activity of an immune checkpoint regulator so that, as a result of the binding to the checkpoint regulator or its ligand, signaling through the checkpoint regulator receptor is blocked or inhibited. By inhibiting this signaling, immune-suppression can be reversed so that T cell immunity against cancer cells can be re-established or enhanced. Immune checkpoint regulator antagonists include antibody fragments, peptide inhibitors, dominant negative peptides and small molecule drugs, either in isolated forms or as part of a fusion protein or conjugate. Example targets of checkpoint regulator antagonists include PD1, POL1, CTLA4, LAG3, TIM-3, TIGIT, VISTA.
The term “immune checkpoint binding agonist” and “immune checkpoint agonist” refer to a class of agents that stimulate the activity of an immune checkpoint regulator so that, as a result of the binding to the checkpoint regulator or its ligand, signaling through the checkpoint regulator receptor is stimulated. By stimulating this signaling, T cell immunity against cancer cells can be re-established or enhanced. The targets of checkpoint regulator agonists include members of the tumor necrosis factor (TNF) receptor superfamily, such as CD27, CD40, OX40 (CD 134), glucocorticoid-induced TNFR family-related protein (GITR), and 4-1BB (CD137) and their ligands. Additional targets of checkpoint regulator agonists belong to the B7-CD28 superfamily, including CD28 and ICOS.
In any of the embodiments above, one or more cancer therapies, e.g., chemotherapy, radiation therapy, immunotherapy, surgery, or hormone therapy can be co-administered further with an antibody of the invention.
In one embodiment, the chemotherapeutic reagent is an alkylating agent: nitrogen mustards, nitrosoureas, tetrazines, aziridines, cisplatins and derivatives, and non-classical alkylating agents. Nitrogen mustards include mechlorethamine, cyclophosphamide, melphalan, chlorambucil, ifosfamide and busulfan. Nitrosoureas include N-Nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU) and semustine (MeCCNU), fotemustine and streptozotocin. Tetrazines include dacarbazine, mitozolomide and temozolomide. Aziridines include thiotepa, mytomycin and diaziquone (AZQ). Cisplatin and derivatives include cisplatin, carboplatin and oxaliplatin. In one embodiment the chemotherapeutic reagent is an anti-metabolites: the anti-folates (e.g., methotrexate), fluoropyrimidines (e.g., fluorouracil and capecitabine), deoxynucleoside analogues and thiopurines. In another embodiment the chemoptheraputic reagent is an anti-microtubule agent such as vinca alkaloids (e.g., vincristine and vinblastine) and taxanes (e.g., paclitaxel and docetaxel). In another embodiment the chemotherapeutic reagent is a topoisomerase inhibitor or a cytotoxic antibiotic such as doxorubicin, mitoxantrone, bleomycin, actinomycin, and mitomycin.
The contacting of the patient with the antibody or antibody fragment, can be by administering the antibody to the patient intravenously, intraperitoneally, intramuscularly, intratumorally, or intradermally. In some embodiments the antibody is co-administered with a cancer therapy agent.
The term “formulation” as used herein refers to the antibodies disclosed herein and excipients combined together which can be administered and has the ability to bind to the corresponding receptors and initiate a signal transduction pathway resulting in the desired activity. The formulation can optionally comprise other agents.
All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are to be understood as approximations in accordance with common practice in the art. When used herein, the term “about” may connote variation (+) or (−) 1%, 5% or 10% of the stated amount, as appropriate given the context. It is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.
Many known and useful compounds and the like can be found in Remington's Pharmaceutical Sciences (13th Ed), Mack Publishing Company, Easton, PA—a standard reference for various types of administration. As used herein, the term “formulation(s)” means a combination of at least one active ingredient with one or more other ingredient, also commonly referred to as excipients, which may be independently active or inactive. The term “formulation” may or may not refer to a pharmaceutically acceptable composition for administration to humans or animals and may include compositions that are useful intermediates for storage or research purposes.
As the patients and subjects of the invention method are, in addition to humans, veterinary subjects, formulations suitable for these subjects are also appropriate. Such subjects include livestock and pets as well as sports animals such as horses, greyhounds, and the like.
DETAILED DESCRIPTION
The DLL3 gene provides instructions for making a protein that helps control the Notch pathway, an important pathway in embryonic development. DLL3 is usually an intracellular protein but it is also expressed on the surface of cancer cells. DLL3 is expressed normally on the inside of cells and at low levels on normal tissues. However, lung tumor cells overexpress the gene and cell surface DLL3 levels are increased. Recent studies have reported that DLL3 is also expressed in other tumor types of neuroendocrine origin, including melanoma, glioblastoma multiforme, small cell bladder cancer, metastatic castration-resistant prostate cancer, and neuroendocrine lung tumors.
CD (cluster of differentiation) molecules (e.g. CD3, CD28 and CD137) CD3, CD28 and CD137 are receptors present on T cells. T cells can be activated by antigen-presenting cells via CD3, CD28 and CD137. Two parallel therapeutic strategies are pursued for activating or engaging T cells to kill tumor cells.
Embodiments of the invention include methods of diagnosing, prognosing, treating, monitoring and preventing cancer, including refractory cancer, using anti-DLL3 antibodies and antibody conjugates, pharmaceutical compositions thereof, and articles of manufacture. More specifically, the invention is directed to bispecific or trispecific molecules that bind to DLL3 and activate CD (cluster of differentiation) molecules (e.g. CD3, CD28 and CD137).
Embodiments of the invention also include methods of diagnosing, prognosing, treating, monitoring and preventing cancer, including refractory cancer, using anti-MUC17 antibodies and antibody conjugates, pharmaceutical compositions thereof, and articles of manufacture. More specifically, the invention is directed to bispecific or trispecific molecules that bind to MUC17 and activate CD (cluster of differentiation) molecules (e.g. CD3, CD28 and CD137).
Embodiments of the invention also include methods of diagnosing, prognosing, treating, monitoring and preventing cancer, including refractory cancer, using anti-CLDN18.2 antibodies and antibody conjugates, pharmaceutical compositions thereof, and articles of manufacture. More specifically, the invention is directed to trispecific molecules that bind to CLDN18.2 and activate CD (cluster of differentiation) molecules (e.g. CD3, CD28 and CD137).
DLL3 T-Cell Engagers
Embodiments of the invention include bispecific and trispecific monoclonal antibodies (BSMAbs and TSMAbs, respectively). The single chain variable fragment (scFv) of a first antibody can be joined with the antigen binding fragment (Fab) of a second antibody. For example, the scFv portion of an antibody against CD3 can be linked to a Fab portion of an antibody against DLL3. They can be joined with 4×GKPGS (SEQ ID NO: 508) or 4×G4S (SEQ ID NO: 509) linkers. The IgG1 Fc can be heterodimerized with charge pair or “knob into hole” mutations or charge pair mutations. The Fc effector function can be minimized through the introduction of N297A/G mutations or LLP mutations. The combination can bring an effector cell (T-cell or NK cell) into the proximity of the tumor cell to enhance antitumor effect.
FIGS. 1A-1R, 8A-8C and 9A-9F depict several formats for trispecific molecules of the invention. The DLL3×CD3 trispecific molecules can activate T cell cytotoxicity against DLL3 expressing CHO cells or NCI-H82 tumor cells. This is exemplified by the release of LDH upon cell death, as well as the upregulation of CD25 on the T cells. When combined with DLL3×CD28 or DLL3×CD137 trispecific molecules, the T cells are further activated, proliferate, and release IFN gamma and IL-2.
CD28 signaling is essential for the activity of anti-PD1 and anti-PDL1 antibodies, thus co-dosing the DLL3×CD28 trispecific molecules with anti-PD1 and anti-PDL1 can improve the responses to the inhibition of the PD1/PDL1 pathway. CD137 is highly expressed on exhausted T cells, which cannot be stimulated by PD1/PDL1 stimulation alone. However, DLL3×CD137 or DLL3×CD28×CD137 stimulation combined with PD-1 blockade results in robust antitumor immunity.
FIG. 1A-1R depict trispecific molecules with combinations of an anti-DLL3 with anti-CD3, anti-CD28 and anti-CD137 in the Scfv-Scfv-Fc×Fab-Fc configuration. FIG. 1A-1F depict trispecific with the configuration scfv-DLL3scfv-Fc×Fab-Fc. FIG. 1G-1L depict trispecific molecules with the configuration DLL3scfv-scfv-Fc×Fab-Fc. FIG. 1M-1R depict depict trispecific molecules with the configuration scfv-scfv-Fc×DLL3Fab-Fc.
FIG. 8A-8C depict bispecific and trispecific molecules with two CD137 Fab fragments. FIG. 8A depicts an anti-CD137, anti-DLL3, anti-DLL3 (Fab-Fc-Scfv) molecule. FIG. 8B depicts an anti-CD137, anti-DLL3, anti-CD3 (Fab-Fc-Scfv×Fab-Fc-Scfv) molecule. FIG. 8C depicts an anti-CD137, anti-DLL3, anti-CD3 (Fab-Fc-Scfv-Scfv×Fab-Fc) molecule.
FIG. 9A-9F depict trispecific molecules with two CD137 fragments. FIG. 9A depicts an anti-DLL3, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc-Scfv×Fc-Scfv) molecule. FIG. 9B depicts an anti-DLL3, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc×Scfv-Fc-Scfv) molecule. FIG. 9C depicts an anti-CD137, anti-CD137, anti-DLL3 (scfv-scfv-Fc×Fab-Fc) molecule. FIG. 9D depicts an anti-DLL3, anti-CD137, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule. FIG. 9E depicts an anti-DLL3, anti-CD3, anti-DLL3, anti-CD137 (Fab-Fc-Scfv×Scfv-Fc-Scfv) molecule. FIG. 9F depicts an anti-DLL3, anti-CD3, anti-CD137, anti-CD137 (Fab-Fc-Scfv-Scfv×Scfv-Fc) molecule.
MUC17 T-Cell Engagers
FIGS. 10A-10R, 18A-18D and 19A-19I depict several formats for trispecific molecules of the invention.
FIG. 10A-10R depict trispecific molecules with combinations of an anti-MUC17 with anti-CD3, anti-CD28 and anti-CD137 in the Scfv-scfv-Fc×Fab-Fc configuration. FIG. 10A-10F depict trispecific with the configuration scfv-MUC17scfv-Fc×Fab-Fc. FIG. 1G-1L depict trispecific molecules with the configuration MUC17scfv-scfv-Fc×Fab-Fc. FIG. 1M-1R depict depict trispecific molecules with the configuration scfv-scfv-Fc×MUC17Fab-Fc.
FIG. 18A-18D depict Muc17×CD137 and DLL3×CD137 bispecific molecules
FIG. 19A-19C depict trispecific molecules with two CD137 Fab fragments. FIG. 19A depicts an anti-CD137, anti-MUC17, anti-MUC17 (Fab-Fc-Scfv-Scfv) molecule. FIG. 19B depicts an anti-CD137, anti-MUC17, anti-CD3 (Fab-Fc-Scfv×Fab-Fc-Scfv) molecule. FIG. 19C depicts an anti-CD137, anti-MUC17, anti-CD3 (Fab-Fc-Scfv-Scfv×Fab-Fc) molecule. FIG. 19D-19I depict bispecific molecules and trispecific molecules with two CD137 scfv fragments. FIG. 19D depicts an, anti-anti-CD137, anti-CD137, anti-MUC17 (scfv-scfv-Fc×Fab-Fc) molecule. FIG. 19E depicts an anti-MUC17, anti-CD137, anti-CD137 (scfv-scfv-Fc×Fab-Fc) molecule. FIG. 19F depicts an anti-MUC17, anti-CD137, CD3, CD137 (Fab-Fc-Scfv×Scfv-Fc-Scfv) molecule. FIG. 19G depicts an anti-MUC17, anti-CD137, anti-CD137, CD3 (Fab-Fc-Scfv-Scfv×Scfv-Fc) molecule. FIG. 19H depicts an anti-MUC17, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-FScfv×Fc-Scfv) molecule. FIG. 19I depicts an anti-DLL3, anti-CD3, anti-CD137, anti-MUC17 (scfv-scfv-Fc×Scfv-Fc-Scfv) molecule.
In an embodiment, the trispecific molecule can be co-administered or combined with an antagonist such as PD1, PDL1, TIGIT, LAG3, TIM3, VISTA or CTLA4. Alternatively, the trispecific molecule can be co-administered or combined with a bispecific antagonist such as PD1×TIGIT, LAG3×TIGIT, PD1×LAG3, PD1×TIM3 or VEGF×TGFBR2. The trispecific molecule can also be co-administered or combined with an agonist such as CD40, GITR, CD27, OX40 or 4-1BB.
CLDN18.2 T-Cell Engagers
Embodiments of the invention include bispecific and trispecific monoclonal antibodies (BSMAbs and TSMAbs, respectively). The single chain variable fragment (scFv) of a first antibody can be joined with the antigen binding fragment (Fab) of a second antibody. For example, the scFv portion of an antibody against CD3 can be linked to a Fab portion of an antibody against CLDN8.2. They can be joined with 4×GKPGS or 4×G4S linkers. The IgG1 Fc can be heterodimerized with charge pair or “knob into hole” mutations or charge pair mutations. The Fc effector function can be minimized through the introduction of N297A/G mutations or LLP mutations. The combination can bring an effector cell (T-cell or NK cell) into the proximity of the tumor cell to enhance antitumor effect.
CD28 signaling is essential for the activity of anti-PD1 and anti-PDL1 antibodies, thus co-dosing the CLDN18.2×CD28 trispecific molecules with anti-PD1 and anti-PDL1 can improve the responses to the inhibition of the PD1/PDL1 pathway. CD137 is highly expressed on exhausted T cells, which cannot be stimulated by PD1/PDL1 stimulation alone. However, CLDN18.2×CD137 or CLDN18.2×CD28×CD137 stimulation combined with PD-1 blockade results in robust antitumor immunity.
FIGS. 20A-20R and 25A-25I depict several formats for trispecific molecules of the invention. The CLDN18.2×CD3 trispecific molecules can activate T cell cytotoxicity against CLDN18.2 expressing CHO cells or NCI-H82 tumor cells. This is exemplified by the release of LDH upon cell death, as well as the upregulation of CD25 on the T cells. When combined with CLDN18.2×CD28 or CLDN18.2×CD137 trispecific molecules, the T cells are further activated, proliferate, and release IFN gamma and IL-2.
FIG. 20A-20R depict trispecific molecules with combinations of an anti-CLDN18.2 with anti-CD3, anti-CD28 and anti-CD137 in the Scfv-scfv-Fc×Fab-Fc configuration. FIG. 20A-20F depict trispecific with the configuration scfv-CLDn18.1scfv-Fc×Fab-Fc. FIG. 20G-20L depict trispecific molecules with the configuration CLDN18.2scfv-scfv-Fc×Fab-Fc. FIG. 20M-20R depict depict trispecific molecules with the configuration scfv-scfv-Fc×CLDN18.2Fab-Fc.
FIG. 25A-25C depict CLDN18.2 bispecific and trispecific molecules with two CD137 Fab fragments.
FIG. 25A depicts an anti-CD137, anti-CLDN18.2, anti-CLDN (Fab-Fc-Scfv) molecule. FIG. 25B depicts an anti-CD137, anti-CLDN18.2, anti-CD3 (Fab-Fc-Scfv×Fab-Fc-Scfv) molecule. FIGS. 25D and 25E depict trispecific molecules with two CD137 fragments.
FIG. 25D depicts an anti-CD137, anti-CD137, anti-CLDN18.2 bispecific molecule (Scfv-Scfv-Fc×Fab-Fc). Similarly, FIG. 25E depicts an anti-CLDN18.2, anti-CD137, anti-CD137, trispecific molecule (Scfv-Scfv-Fc×Fab-Fc)
FIG. 25F-25I depict trispecific molecules with two CD137 scfv fragments. FIG. 25F depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (Fab-Fc-Scfv×ScfvFc-Scfv) molecule. FIG. 25G depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (Fab-Fc-Scfv-Scfv×Scfv-Fc) molecule. FIG. 25H depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc-Scfv×Fc-Scfv) molecule. FIG. 25I depicts an anti-CLDN18.2, anti-CD3, anti-CD137, anti-CD137 (scfv-scfv-Fc×Scfv-Fc-Scfv) molecule.
In an embodiment, the trispecific molecule can be co-administered or combined with an antagonist such as PD1, PDL1, TIGIT, LAG3, TIM3, VISTA or CTLA4. Alternatively, the trispecific molecule can be co-administered or combined with a bispecific antagonist such as PD1×TIGIT, LAG3×TIGIT, PD1×LAG3, PD1×TIM3 or VEGF×TGFBR2. The trispecific molecule can also be co-administered or combined with an agonist such as CD40, GITR, CD27, OX40 or 4-1BB.
Methods from Producing Trispecific T-Cell Engagers
Another aspect relates to a method for producing a trispecific antibody comprising culturing a cell transiently or stably expressing one or more constructs encoding one or more polypeptide chains in the trispecific antibody; and purifying the trispecific antibody from the cultured cells. Any cell capable of producing a functional trispecific antibody can be used. In preferred embodiments, the trispecific antibody-expressing cell is of eukaryotic or mammalian origin, preferably a human cell or Chinese hamster cell. Cells from various tissue cell types may be used to express the trispecific antibodies. In other embodiments, the cell is a yeast cell, an insect cell or a bacterial cell. Preferably, the trispecific antibody-producing cell is stably transformed with a vector expressing the trispecific antibody.
One or more expression vectors encoding the antibody heavy or light chains can be introduced into a cell by any conventional method, such as by naked DNA technique, cationic lipid-mediated transfection, polymer-mediated transfection, peptide-mediated transfection, virus-mediated infection, physical or chemical agents or treatments, electroporation, etc. In addition, cells may be transfected with one or more expression vectors for expressing the trispecific antibody along with a selectable marker facilitating selection of stably transformed clones expressing the trispecific antibody. The antibodies produced by such cells may be collected and/or purified according to techniques known in the art, such as by centrifugation, chromatography, etc.
Examples of suitable selectable markers for mammalian cells include dihydrofolate reductase (DHFR), thymidine kinase, neomycin, neomycin analog G418, hydromycin, zeocin, blasticidin, and puromycin. When such selectable markers are successfully transferred into a mammalian host cell, the transformed mammalian host cell can survive if placed under selective pressure. There are two widely used distinct categories of selective regimes. The first category is based on a cell's metabolism and the use of a mutant cell line which lacks the ability to grow independent of a supplemented media. Two examples are CHO DHFR cells and mouse LTK cells. These cells lack the ability to grow without the addition of such nutrients as thymidine or hypoxanthine. Because these cells lack certain genes necessary for a complete nucleotide synthesis pathway, they cannot survive unless the missing nucleotides are provided in a supplemented media. An alternative to supplementing the media is to introduce an intact DHFR or TK gene into cells lacking the respective genes, thus altering their growth requirements. Individual cells which were not transformed with the DHFR or TK gene will not be capable of survival in non-supplemented media.
The second category is dominant selection which refers to a selection scheme used in any cell type and does not require the use of a mutant cell line. These schemes typically use a drug to arrest growth of a host cell. Those cells which have a novel gene would express a protein conveying drug resistance and would survive the selection. Examples of such dominant selection use the drugs neomycin, mycophenolic acid, or hygromycin. The three examples employ bacterial genes under eukaryotic control to convey resistance to the appropriate drug G418 or neomycin (geneticin), xgpt (mycophenolic acid) or hygromycin, respectively. Others include the neomycin analog G418 and puromycin.
Exemplary trispecific antibody-expressing cells include human Jurkat, human embryonic kidney (HEK) 293, Chinese hamster ovary (CHO) cells, mouse WEHI fibrosarcoma cells, as well as unicellular protozoan species, such as Leishmania tarentolae. In addition, stably transformed, antibody producing cell lines may be produced using primary cells immortalized with c-myc or other immortalizing agents.
In some embodiments, the cell lines express at least 1 mg, at least 2 mg, at least 5 mg, at least 10 mg, at least 20 mg, at least 50 mg, at least 100 mg, at least 500 mg, at least 1 gram, at least 2 grams, at least 4 grams, or at least 10 grams of the trispecific antibody/liter of culture. Trispecific antibodies can be isolated from trispecific antibody expressing cells following culture and maintenance in any appropriate culture medium, such as RPMI, DMEM, and AIM VO. The trispecific antibodies can be purified using conventional protein purification methodologies (e.g., affinity purification, chromatography, etc.), including the use of Protein-A or Protein-G immunoaffinity purification. In some embodiments, trispecific antibodies are engineered for secretion into culture supernatants for isolation therefrom.
Methods of Treatment
Another aspect of the present application relates to a method for treating a cell proliferative disorder. The method comprises administering to a subject in need thereof an effective amount of a trispecific antibody according to the present disclosure. In another aspect, a method for treating a cell proliferative disorder comprises administering to a subject in need thereof an effective amount of one or more expression vectors expressing a trispecific antibody according to the present disclosure.
Any suitable route or mode of administration can be employed for providing the patient with a therapeutically or prophylactically effective dose of the trispecific antibody. Exemplary routes or modes of administration include parenteral {e.g., intravenous, intraarterial, intramuscular, subcutaneous, intratumoral), oral, topical (nasal, transdermal, intradermal or intraocular), mucosal {e.g., nasal, sublingual, buccal, rectal, vaginal), inhalation, intralymphatic, intraspinal, intracranial, intraperitoneal, intratracheal, intravesical, intrathecal, enteral, intrapulmonary, intralymphatic, intracavital, intraorbital, intracapsular and transurethral, as well as local delivery by catheter or stent.
A pharmaceutical composition comprising a trispecific antibody in accordance with the present disclosure can be formulated in any pharmaceutically acceptable carrier(s) or excipient(s). As used herein, the term “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Pharmaceutical compositions can include suitable solid or gel phase carriers or excipients. Exemplary carriers or excipients include calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols. Exemplary pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like, as well as combinations thereof. In many cases it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable carriers can further comprise minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the therapeutic agents.
The trispecific antibody can be incorporated into a pharmaceutical composition suitable for parenteral administration. Suitable buffers include but are not limited to, sodium succinate, sodium citrate, sodium phosphate or potassium phosphate. Sodium chloride can be used to modify the toxicity of the solution at a concentration of 0-300 mM (optimally 150 mM for a liquid dosage form). Cryoprotectants can be included for a lyophilized dosage form, principally 0-10% sucrose (optimally 0.5-1.0%). Other suitable cryoprotectants include trehalose and lactose. Bulking agents can be included for a lyophilized dosage form, principally 1-10% mannitol (optimally 2-4%). Stabilizers can be used in both liquid and lyophilized dosage forms, principally 1-50 mM L-Methionine (optimally 5-10 mM). Other suitable bulking agents include glycine, arginine, can be included as 0-0.05%>polysorbate-80 (optimally 0.005-0.01%). Additional surfactants include but are not limited to polysorbate 20 and BRIJ surfactants.
Therapeutic trispecific antibody preparations can be lyophilized and stored as sterile powders, preferably under vacuum, and then reconstituted in bacteriostatic water (containing, for example, benzyl alcohol preservative) or in sterile water prior to injection. Pharmaceutical compositions can be formulated for parenteral administration by injection e.g., by bolus injection or continuous infusion.
The therapeutic agents in the pharmaceutical compositions may be formulated in a “therapeutically effective amount” or a “prophylactically effective amount”. A “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the recombinant vector may vary depending on the condition to be treated, the severity and course of the condition, the mode of administration, whether the antibody or agent is administered for preventive or therapeutic purposes, the bioavailability of the particular agent(s), the ability of the trispecific antibody to elicit a desired response in the individual, previous therapy, the age, weight and sex of the patient, the patient's clinical history and response to the antibody, the type of the trispecific antibody used, discretion of the attending physician, etc. A therapeutically effective amount is also one in which any toxic or detrimental effects of the recombinant vector is outweighed by the therapeutically beneficial effects. A “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result.
Preferably, the polypeptide domains in the trispecific antibody are derived from the same host in which they are to be administered in order to reduce inflammatory responses against the administered therapeutic agents.
The trispecific antibody is suitably administered to the patent at one time or over a series of treatments and may be administered to the patient at any time from diagnosis onwards. The trispecific antibody may be administered as the sole treatment or in conjunction with other drugs or therapies useful in treating the condition in question.
As a general proposition, a therapeutically effective amount or prophylactically effective amount of the trispecific antibody will be administered in a range from about 1 ng/kg body weight/day to about 100 mg/kg body weight/day whether by one or more administrations. In a particular embodiment, each trispecific antibody is administered in the range of from about 1 ng/kg body weight/day to about 10 mg/kg body weight/day, about 1 ng/kg body weight/day to about 1 mg/kg body weight/day, about 1 ng/kg body weight/day to about 100 g/kg body weight/day, about 1 ng/kg body weight/day to about 10 g/kg body weight/day, about 1 ng/kg body weight/day to about 1 g/kg body weight/day, about 1 ng/kg body weight/day to about 100 ng/kg body weight/day, about 1 ng/kg body weight/day to about 10 ng/kg body weight/day, about 10 ng/kg body weight/day to about 100 mg/kg body weight/day, about 10 ng/kg body weight/day to about 10 mg/kg body weight/day, about 10 ng/kg body weight/day to about 1 mg/kg body weight/day, about 10 ng/kg body weight/day to about 100 g/kg body weight/day, about 10 ng/kg body weight/day to about 10 mg/kg body weight/day, about 10 ng/kg body weight/day to about 1 mg/kg body weight/day, 10 ng/kg body weight/day to about 100 ng/kg body weight/day, about 100 ng/kg body weight/day to about 100 mg/kg body weight/day, about 100 ng/kg body weight/day to about 10 mg/kg body weight/day, about 100 ng/kg body weight/day to about 1 mg/kg body weight/day, about 100 ng/kg body weight/day to about 100 mg/kg body weight/day, about 100 ng/kg body weight/day to about 10 mg/kg body weight/day, about 100 ng/kg body weight/day to about 1 mg/kg body weight/day, about 1 mg/kg body weight/day to about 100 mg/kg body weight/day, about 1 mg/kg body weight/day to about 10 mg/kg body weight/day, about 1 mg/kg body weight/day to about 1 mg/kg body weight/day, about 1 mg/kg body weight/day to about 100 mg/kg body weight/day, about 1 mg/kg body weight/day to about 10 mg/kg body weight/day, about 10 mg/kg body weight/day to about 100 mg/kg body weight/day, about 10 mg/kg body weight/day to about 10 mg/kg body weight/day, about 10 mg/kg body weight/day to about 1 mg/kg body weight/day, about 10 mg/kg body weight/day to about 100 mg/kg body weight/day, about 100 mg/kg body weight/day to about 100 mg/kg body weight/day, about 100 mg/kg body weight/day to about 10 mg/kg body weight/day, about 100 mg/kg body weight/day to about 1 mg/kg body weight/day, about 1 mg/kg body weight/day to about 100 mg/kg body weight/day, about 1 mg/kg body weight/day to about 10 mg/kg body weight/day, about 10 mg/kg body weight/day to about 100 mg/kg body weight/day.
In other embodiments, the trispecific antibody is administered at a dose of 500 g to 20 g every three days, or 25 mg/kg body weight every three days.
In other embodiments, each trispecific antibody is administered in the range of about 10 ng to about 100 ng per individual administration, about 10 ng to about 1 g per individual administration, about 10 ng to about 10 g per individual administration, about 10 ng to about 100 mg per individual administration, about 10 ng to about 1 mg per individual administration, about 10 ng to about 10 mg per individual administration, about 10 ng to about 100 mg per individual administration, about 10 ng to about 1000 mg per injection, about 10 ng to about 10,000 mg per individual administration, about 100 ng to about 1 mg per individual administration, about 100 ng to about 10 mg per individual administration, about 100 ng to about 100 mg per individual administration, about 100 ng to about 1 mg per individual administration, about 100 ng to about 10 mg per individual administration, about 100 ng to about 100 mg per individual administration, about 100 ng to about 1000 mg per injection, about 100 ng to about 10,000 mg per individual administration, about 1 mg to about 10 mg per individual administration, about 1 mg to about 100 mg per individual administration, about 1 mg to about 1 mg per individual administration, about 1 mg to about 10 mg per individual administration, about 1 mg to about 100 mg per individual administration, about 1 mg to about 1000 mg per injection, about 1 mg to about 10,000 mg per individual administration, about 10 mg to about 100 mg per individual administration, about 10 mg to about 1 mg per individual administration, about 10 mg to about 10 mg per individual administration, about 10 mg to about 100 mg per individual administration, about 10 mg to about 1000 mg per injection, about 10 mg to about 10,000 mg per individual administration, about 100 mg to about 1 mg per individual administration, about 100 mg to about 10 mg per individual administration, about 100 mg to about 100 mg per individual administration, about 100 mg to about 1000 mg per injection, about 100 mg to about 10,000 mg per individual administration, about 1 mg to about 10 mg per individual administration, about 1 mg to about 100 mg per individual administration, about 1 mg to about 1000 mg per injection, about 1 mg to about 10,000 mg per individual administration, about 10 mg to about 100 mg per individual administration, about 10 mg to about 1000 mg per injection, about 10 mg to about 10,000 mg per individual administration, about 100 mg to about 1000 mg per injection, about 100 mg to about 10,000 mg per individual administration and about 1000 mg to about 10,000 mg per individual administration. The trispecific antibody may be administered daily, every 2, 3, 4, 5, 6 or 7 days, or every 1, 2, 3 or 4 weeks.
In other particular embodiments, the amount of the trispecific antibody may be administered at a dose of about 0.0006 mg/day, 0.001 mg/day, 0.003 mg/day, 0.006 mg/day, 0.01 mg/day, 0.03 mg/day, 0.06 mg/day, 0.1 mg/day, 0.3 mg/day, 0.6 mg/day, 1 mg/day, 3 mg/day, 6 mg/day, 10 mg/day, 30 mg/day, 60 mg/day, 100 mg/day, 300 mg/day, 600 mg/day, 1000 mg/day, 2000 mg/day, 5000 mg/day or 10,000 mg/day. As expected, the dosage will be dependent on the condition, size, age and condition of the patient.
In certain embodiments, the coding sequences for a trispecific antibody are incorporated into a suitable expression vector (e.g., viral or non-viral vector) for expressing an effective amount of the trispecific antibody in patient with a cell proliferative disorder. In certain embodiments comprising administration of e.g., one or more recombinant AAV (rAAV) viruses, the pharmaceutical composition may comprise the rAAVs in an amount comprising at least 1010, at least 1011, at least 1012, at least 1013, or at least 1014 genome copies (GC) or recombinant viral particles per kg, or any range thereof. In certain embodiments, the pharmaceutical composition comprises an effective amount of the recombinant virus, such as rAAV, in an amount comprising at least 1010, at least 1011, at least 1012, at least 1013, at least 1014, at least 1015 genome copies or recombinant viral particles genome copies per subject, or any range thereof.
Dosages can be tested in several art-accepted animal models suitable for any particular cell proliferative disorder.
Delivery methodologies may also include the use of polycationic condensed DNA linked or unlinked to killed viruses, ligand linked DNA, liposomes, eukaryotic cell delivery vehicles cells, deposition of photopolymerized hydrogel materials, use of a handheld gene transfer particle gun, ionizing radiation, nucleic charge neutralization or fusion with cell membranes, particle mediated gene transfer and the like.
In other aspects of this embodiment, a pharmaceutical composition compound disclosed herein reduces the size of a tumor by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In yet other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the size of a tumor from, e.g., about 5% to about 100%, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.
A pharmaceutical composition disclosed herein is in an amount sufficient to allow customary administration to an individual. In aspects of this embodiment, a pharmaceutical composition disclosed herein may be, e.g., at least 5 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mg of a pharmaceutical composition. In other aspects of this embodiment, a pharmaceutical composition disclosed herein may be, e.g., at least 5 mg, at least 10 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, at least 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, at least 1,000 mg, at least 1,100 mg, at least 1,200 mg, at least 1,300 mg, at least 1,400 mg, or at least 1,500 mg of a pharmaceutical composition. In yet other aspects of this embodiment, a pharmaceutical composition disclosed herein may be in the range of, e.g., about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 50 mg to about 150 mg, about 100 mg to about 250 mg, about 150 mg to about 350 mg, about 250 mg to about 500 mg, about 350 mg to about 600 mg, about 500 mg to about 750 mg, about 600 mg to about 900 mg, about 750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, or about 1,000 mg to about 1,500 mg. In still other aspects of this embodiment, a pharmaceutical composition disclosed herein may be in the range of, e.g., about 10 mg to about 250 mg, about 10 mg to about 500 mg, about 10 mg to about 750 mg, about 10 mg to about 1,000 mg, about 10 mg to about 1,500 mg, about 50 mg to about 250 mg, about 50 mg to about 500 mg, about 50 mg to about 750 mg, about 50 mg to about 1,000 mg, about 50 mg to about 1,500 mg, about 100 mg to about 250 mg, about 100 mg to about 500 mg, about 100 mg to about 750 mg, about 100 mg to about 1,000 mg, about 100 mg to about 1,500 mg, about 200 mg to about 500 mg, about 200 mg to about 750 mg, about 200 mg to about 1,000 mg, about 200 mg to about 1,500 mg, about 5 mg to about 1,500 mg, about 5 mg to about 1,000 mg, or about 5 mg to about 250 mg.
A pharmaceutical composition disclosed herein may comprise a solvent, emulsion or other diluent in an amount sufficient to dissolve a pharmaceutical composition disclosed herein. In other aspects of this embodiment, a pharmaceutical composition disclosed herein may comprise a solvent, emulsion or a diluent in an amount of, e.g., less than about 90% (v/v), less than about 80% (v/v), less than about 70% (v/v), less than about 65% (v/v), less than about 60% (v/v), less than about 55% (v/v), less than about 50% (v/v), less than about 45% (v/v), less than about 40% (v/v), less than about 35% (v/v), less than about 30% (v/v), less than about 25% (v/v), less than about 20% (v/v), less than about 15% (v/v), less than about 10% (v/v), less than about 5% (v/v), or less than about 1% (v/v). In other aspects of this embodiment, a pharmaceutical composition disclosed herein may comprise a solvent, emulsion or other diluent in an amount in a range of, e.g., about 1% (v/v) to 90% (v/v), about 1% (v/v) to 70% (v/v), about 1% (v/v) to 60% (v/v), about 1% (v/v) to 50% (v/v), about 1% (v/v) to 40% (v/v), about 1% (v/v) to 30% (v/v), about 1% (v/v) to 20% (v/v), about 1% (v/v) to 10% (v/v), about 2% (v/v) to 50% (v/v), about 2% (v/v) to 40% (v/v), about 2% (v/v) to 30% (v/v), about 2% (v/v) to 20% (v/v), about 2% (v/v) to 10% (v/v), about 4% (v/v) to 50% (v/v), about 4% (v/v) to 40% (v/v), about 4% (v/v) to 30% (v/v), about 4% (v/v) to 20% (v/v), about 4% (v/v) to 10% (v/v), about 6% (v/v) to 50% (v/v), about 6% (v/v) to 40% (v/v), about 6% (v/v) to 30% (v/v), about 6% (v/v) to 20% (v/v), about 6% (v/v) to 10% (v/v), about 8% (v/v) to 50% (v/v), about 8% (v/v) to 40% (v/v), about 8% (v/v) to 30% (v/v), about 8% (v/v) to 20% (v/v), about 8% (v/v) to 15% (v/v), or about 8% (v/v) to 12% (v/v).
The final concentration of a pharmaceutical composition disclosed herein in a pharmaceutical composition disclosed herein may be of any concentration desired. In an aspect of this embodiment, the final concentration of a pharmaceutical composition in a pharmaceutical composition may be a therapeutically effective amount. In other aspects of this embodiment, the final concentration of a pharmaceutical composition in a pharmaceutical composition may be, e.g., at least 0.00001 mg/mL, at least 0.0001 mg/mL, at least 0.001 mg/mL, at least 0.01 mg/mL, at least 0.1 mg/mL, at least 1 mg/mL, at least 10 mg/mL, at least 25 mg/mL, at least 50 mg/mL, at least 100 mg/mL, at least 200 mg/mL or at least 500 mg/mL. In other aspects of this embodiment, the final concentration of a pharmaceutical composition in a pharmaceutical composition may be in a range of, e.g., about 0.00001 mg/mL to about 3,000 mg/mL, about 0.0001 mg/mL to about 3,000 mg/mL, about 0.01 mg/mL to about 3,000 mg/mL, about 0.1 mg/mL to about 3,000 mg/mL, about 1 mg/mL to about 3,000 mg/mL, about 250 mg/mL to about 3,000 mg/mL, about 500 mg/mL to about 3,000 mg/mL, about 750 mg/mL to about 3,000 mg/mL, about 1,000 mg/mL to about 3,000 mg/mL, about 100 mg/mL to about 2,000 mg/mL, about 250 mg/mL to about 2,000 mg/mL, about 500 mg/mL to about 2,000 mg/mL, about 750 mg/mL to about 2,000 mg/mL, about 1,000 mg/mL to about 2,000 mg/mL, about 100 mg/mL to about 1,500 mg/mL, about 250 mg/mL to about 1,500 mg/mL, about 500 mg/mL to about 1,500 mg/mL, about 750 mg/mL to about 1,500 mg/mL, about 1,000 mg/mL to about 1,500 mg/mL, about 100 mg/mL to about 1,200 mg/mL, about 250 mg/mL to about 1,200 mg/mL, about 500 mg/mL to about 1,200 mg/mL, about 750 mg/mL to about 1,200 mg/mL, about 1,000 mg/mL to about 1,200 mg/mL, about 100 mg/mL to about 1,000 mg/mL, about 250 mg/mL to about 1,000 mg/mL, about 500 mg/mL to about 1,000 mg/mL, about 750 mg/mL to about 1,000 mg/mL, about 100 mg/mL to about 750 mg/mL, about 250 mg/mL to about 750 mg/mL, about 500 mg/mL to about 750 mg/mL, about 100 mg/mL to about 500 mg/mL, about 250 mg/mL to about 500 mg/mL, about 0.00001 mg/mL to about 0.0001 mg/mL, about 0.00001 mg/mL to about 0.001 mg/mL, about 0.00001 mg/mL to about 0.01 mg/mL, about 0.00001 mg/mL to about 0.1 mg/mL, about 0.00001 mg/mL to about 1 mg/mL, about 0.001 mg/mL to about 0.01 mg/mL, about 0.001 mg/mL to about 0.1 mg/mL, about 0.001 mg/mL to about 1 mg/mL, about 0.001 mg/mL to about 10 mg/mL, or about 0.001 mg/mL to about 100 mg/mL.
Aspects of the present specification disclose, in part, treating an individual suffering from cancer. As used herein, the term “treating,” refers to reducing or eliminating in an individual a clinical symptom of cancer; or delaying or preventing in an individual the onset of a clinical symptom of cancer. For example, the term “treating” can mean reducing a symptom of a condition characterized by a cancer, including, but not limited to, tumor size, by, e.g., at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% at least 95%, or at least 100%. The actual symptoms associated with cancer are well known and can be determined by a person of ordinary skill in the art by taking into account factors, including, without limitation, the location of the cancer, the cause of the cancer, the severity of the cancer, and/or the tissue or organ affected by the cancer. Those of skill in the art will know the appropriate symptoms or indicators associated with a specific type of cancer and will know how to determine if an individual is a candidate for treatment as disclosed herein.
In another aspect, a pharmaceutical composition disclosed herein reduces the severity of a symptom of a disorder associated with a cancer. In aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the severity of a symptom of a disorder associated with a cancer by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the severity of a symptom of a disorder associated with a cancer by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.
In aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein reduces a symptom associated with cancer by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 100%. In other aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein reduces a symptom associated with cancer by, e.g., at most 10%, at most 15%, at most 20%, at most 25%, at most 30%, at most 35%, at most 40%, at most 45%, at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, at most 95% or at most 100%. In yet other aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein reduces a symptom associated with cancer by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 20%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.
In yet other aspects of this embodiment, a therapeutically effective amount of a pharmaceutical composition disclosed herein generally is in the range of about 0.001 mg/kg to about 100 mg/kg and administered, for example, every 3, 5, 7, 10 or 14 days. In aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be, e.g., at least 0.001 mg/kg, at least 0.01 mg/kg, at least 0.1 mg/kg, at least 1.0 mg/kg, at least 5.0 mg/kg, at least 10 mg/kg, at least 15 mg/kg, at least 20 mg/kg, at least 25 mg/kg, at least 30 mg/kg, at least 35 mg/kg, at least 40 mg/kg, at least 45 mg/kg, or at least 50 mg/kg and administered, for example, every 3, 5, 7, 10 or 14 days. In other aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be in the range of, e.g., about 0.001 mg/kg to about 10 mg/kg, about 0.001 mg/kg/day to about 15 mg/kg, about 0.001 mg/kg to about 20 mg/kg, about 0.001 mg/kg to about 25 mg/kg, about 0.001 mg/kg to about 30 mg/kg, about 0.001 mg/kg to about 35 mg/kg, about 0.001 mg/kg to about 40 mg/kg, about 0.001 mg/kg to about 45 mg/kg, about 0.001 mg/kg to about 50 mg/kg, about 0.001 mg/kg to about 75 mg/kg, or about 0.001 mg/kg to about 100 mg/kg and administered, for example, every 3, 5, 7, 10 or 14 days. In yet other aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be in the range of, e.g., about 0.01 mg/kg to about 10 mg/kg, about 0.01 mg/kg to about 15 mg/kg, about 0.01 mg/kg to about 20 mg/kg, about 0.01 mg/kg to about 25 mg/kg, about 0.01 mg/kg to about 30 mg/kg, about 0.01 mg/kg to about 35 mg/kg, about 0.01 mg/kg to about 40 mg/kg, about 0.01 mg/kg to about 45 mg/kg, about 0.01 mg/kg to about 50 mg/kg, about 0.01 mg/kg to about 75 mg/kg, or about 0.01 mg/kg to about 100 mg/kg and administered, for example, every 3, 5, 7, 10 or 14 days. In still other aspects of this embodiment, an effective amount of a pharmaceutical composition disclosed herein may be in the range of, e.g., about 0.1 mg/kg to about 10 mg/kg, about 0.1 mg/kg to about 15 mg/kg, about 0.1 mg/kg to about 20 mg/kg, about 0.1 mg/kg to about 25 mg/kg, about 0.1 mg/kg to about 30 mg/kg, about 0.1 mg/kg to about 35 mg/kg, about 0.1 mg/kg to about 40 mg/kg, about 0.1 mg/kg to about 45 mg/kg, about 0.1 mg/kg to about 50 mg/kg, about 0.1 mg/kg to about 75 mg/kg, or about 0.1 mg/kg to about 100 mg/kg and administered, for example, every 3, 5, 7, 10 or 14 days.
Dosing can be single dosage or cumulative (serial dosing), and can be readily determined by one skilled in the art. For instance, treatment of a cancer may comprise a one-time administration of an effective dose of a pharmaceutical composition disclosed herein. Alternatively, treatment of a cancer may comprise multiple administrations of an effective dose of a pharmaceutical composition carried out over a range of time periods, such as, e.g., once daily, twice daily, trice daily, once every few days, or once weekly. The timing of administration can vary from individual to individual, depending upon such factors as the severity of an individual's symptoms. For example, an effective dose of a pharmaceutical composition disclosed herein can be administered to an individual once daily for an indefinite period of time, or until the individual no longer requires therapy. A person of ordinary skill in the art will recognize that the condition of the individual can be monitored throughout the course of treatment and that the effective amount of a pharmaceutical composition disclosed herein that is administered can be adjusted accordingly.
In one embodiment, a cancer therapeutic disclosed herein is capable of reducing the number of cancer cells or tumor size in an individual suffering from a cancer by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% as compared to a patient not receiving the same treatment. In other aspects of this embodiment, a cancer therapeutic is capable of reducing the number of cancer cells or tumor size in an individual suffering from a cancer by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70% as compared to a patient not receiving the same treatment.
In a further embodiment, a cancer therapeutic and its derivatives have half-lives of 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, one month, two months, three months, four months or more.
In an embodiment, the period of administration of a cancer therapeutic is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In a further embodiment, a period of during which administration is stopped is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more.
In aspects of this embodiment, a therapeutically effective amount of a cancer therapeutic disclosed herein reduces or maintains a cancer cell population and/or tumor cell size in an individual by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 100%. In other aspects of this embodiment, a therapeutically effective amount of a cancer therapeutic disclosed herein reduces or maintains a cancer cell population and/or tumor cell size in an individual by, e.g., at most 10%, at most 15%, at most 20%, at most 25%, at most 30%, at most 35%, at most 40%, at most 45%, at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, at most 95% or at most 100%. In yet other aspects of this embodiment, a therapeutically effective amount of a cancer therapeutic disclosed herein reduces or maintains a cancer cell population and/or tumor cell size in an individual by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 20%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%.
A pharmaceutical composition or cancer therapeutic is administered to an individual. An individual is typically a human being, but can be an animal, including, but not limited to, dogs, cats, birds, cattle, horses, sheep, goats, reptiles and other animals, whether domesticated or not. Typically, any individual who is a candidate for treatment is a candidate with some form of cancer, whether the cancer is benign or malignant, a tumor, solid or otherwise, a cancer call not located in a tumor or some other form of cancer. Among the most common types of cancer include, but are not limited to, bladder cancer, breast cancer, colon and rectal cancer, endometrial cancer, kidney cancer, renal cancer, leukemia, lung cancer, melanoma, non-Hodgkins lymphoma, pancreatic cancer, prostate cancer, stomach cancer and thyroid cancer. Pre-operative evaluation typically includes routine history and physical examination in addition to thorough informed consent disclosing all relevant risks and benefits of the procedure.
In one aspect, a pharmaceutical composition disclosed herein reduces a symptom of a disorder associated with a cancer. In aspects of this embodiment, a pharmaceutical composition disclosed herein reduces a symptom of a disorder associated with a cancer by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces a symptom of a disorder associated with a cancer by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.
In another aspect, a pharmaceutical composition disclosed herein reduces the frequency of a symptom of a disorder associated with a cancer incurred over a given time period. In aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the frequency of a symptom of a disorder associated with a cancer incurred over a given time period by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the frequency of a symptom of a disorder associated with a cancer incurred over a given time period by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.







TABLE 1







DLL3 HC and LC Pairs
















VH
VL
HCDR1
HCDR2
HDCR3
LCDR1
LCDR2
LCDR3


Full ab name
Seq ID
seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID





DLL3.3 
72
76
7
14
27
31
40
47


DLL3.4 
73
77
1
11
22
32
39
54


DLL3.26
58
78
6
16
24
33
43
49


DLL3.27
65
78
5
16
24
33
43
49


DLL3.1 
71
79
2
10
22
32
46
55


DLL3.2 
73
79
1
11
22
32
46
55


DLL3.8 
67
80
4
12
23
36
44
48


DLL3.9 
64
81
3
13
28
32
46
54


DLL3.22
57
82
6
15
25
33
43
51


DLL3.23
59
82
6
16
25
33
43
51


DLL3.24
61
82
6
18
25
33
43
51


DLL3.25
63
82
6
16
25
33
43
51


DLL3.15
56
83
6
15
24
34
43
49


DLL3.5 
58
83
6
16
24
34
43
49


DLL3.16
60
83
6
18
24
34
43
49


DLL3.17
62
83
6
16
24
34
43
49


DLL3.18
57
84
6
15
25
37
42
51


DLL3.19
59
84
6
16
25
37
42
51


DLL3.20
61
84
6
18
25
37
42
51


DLL3.21
63
84
6
16
25
37
42
51


DLL3.11
56
85
7
15
24
38
42
49


DLL3.12
58
85
6
16
24
38
42
49


DLL3.13
60
85
6
18
24
38
42
49


DLL3.14
62
85
6
16
24
38
42
49


DLL3.32
70
86
5
16
25
33
43
51


DLL3.33
70
86
5
16
25
33
43
51


DLL3.10
69
87
5
16
24
34
43
49


DLL3.31
69
87
5
16
24
34
43
49


DLL3.29
68
88
5
17
26
34
43
50


DLL3.30
68
88
5
17
26
34
43
50


DLL3.36
68
88
5
17
26
34
43
50


DLL3.34
74
89
8
19
23
35
45
52


DLL3.28
66
90
9
21
29
32
39
54
















TABLE 2







DLL3-CD3












Molecule
Seq ID
seq ID
seq ID


format
Name
chain1
chain2
chain3





DLL3-
3D34C
341
239
—


scFvxCD3-






scFv






DLL3-
3D35C
342
239
—


scFvxCD3-






scFv






DLL3-
3D36C
346
239
—


scFvxCD3-






scFv






DLL3-
3D36D
343
239
—


scFvxCD3-






scFv






DLL3-
3D36I
347
239
—


scFvxCD3-






scFv






DLL3-
3D36K
348
239
—


scFvxCD3-






scFv






DLL3-
3D37C
345
239
—


scFvxCD3-






scFv






DLL3-
3D44I
344
239
—


scFvxCD3-






scFv






DLL3-
3D1
273
290
298


Fab/CD3-scFv






DLL3-
3D10
273
292
296


Fab/CD3-scFv






DLL3-
3D10B
262
292
296


Fab/CD3-scFv






DLL3-
3D10C
263
292
296


Fab/CD3-scFv






DLL3-
3D11B
262
286
310


Fab/CD3-scFv






DLL3-
3D11C
263
286
310


Fab/CD3-scFv






DLL3-
3D12B
262
288
308


Fab/CD3-scFv






DLL3-
3D12C
263
288
308


Fab/CD3-scFv






DLL3-
3D13B
262
289
306


Fab/CD3-scFv






DLL3-
3D34-22C
341
276
302


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D35-22C
342
276
302


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D36-22C
346
276
302


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D37-22C
345
276
302


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D13C
263
289
306


Fab/CD3-scFv






DLL3-
3D45I
347
276
297


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D14B
262
284
312


Fab/CD3-scFv






DLL3-
3D14C
263
284
312


Fab/CD3-scFv






DLL3-
3D15C
263
287
309


Fab/CD3-scFv






DLL3-
3D16C
263
288
307


Fab/CD3-scFv






DLL3-
3D17C
263
289
305


Fab/CD3-scFv






DLL3-
3D18C
263
276
304


Fab/CD3-scFv






DLL3-
3D19C
263
274
304


Fab/CD3-scFv






DLL3-
3D1B
262
290
298


Fab/CD3-scFv






DLL3-
3D1C
263
290
298


Fab/CD3-scFv






DLL3-
3D1I
269
290
298


Fab/CD3-scFv






DLL3-
3D20C
263
278
304


Fab/CD3-scFv






DLL3-
3D21C
263
280
304


Fab/CD3-scFv






DLL3-
3DBM
Benchmark




scFvxCD3-

CD3xDLL3




scFv-Fc-Fc






DLL3-
3D22C
263
276
302


Fab/CD3-scFv






DLL3-
3D22D
264
276
302


Fab/CD3-scFv






DLL3-
3D22I
269
276
302


Fab/CD3-scFv






DLL3-
3D22K
271
276
302


Fab/CD3-scFv






DLL3-
3D23C
263
274
302


Fab/CD3-scFv






DLL3-
3D24C
263
278
302


Fab/CD3-scFv






DLL3-
3D25C
263
280
302


Fab/CD3-scFv






DLL3-
3D26C
263
277
303


Fab/CD3-scFv






DLL3-
3D27C
263
275
303


Fab/CD3-scFv






DLL3-
3D34-16C
341
288
307


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D35-16C
342
288
307


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D36-16C
346
288
307


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D37-16C
345
288
307


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D28C
263
279
303


Fab/CD3-scFv






DLL3-
3D29C
263
281
303


Fab/CD3-scFv






DLL3-
3D30C
263
277
301


Fab/CD3-scFv






DLL3-
3D31C
263
275
301


Fab/CD3-scFv






DLL3-
3D32C
263
279
301


Fab/CD3-scFv






DLL3-
3D33C
263
281
301


Fab/CD3-scFv






DLL3-
3D38I
269
283
297


Fab/CD3-scFv






DLL3-
3D34-1C
341
290
298


scFvxCD3-






scFv/DLL3-Fab






DLL3-
3D35-1C
342
290
298


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D36-1C
346
290
298


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D37-1C
345
290
298


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D39I
269
276
297


Fab/CD3-scFv






DLL3-
3D4
273
291
295


Fab/CD3-scFv






DLL3-
3D40I
269
285
299


Fab/CD3-scFv






DLL3-
3D41I
269
282
300


Fab/CD3-scFv






DLL3-
3D42I
269
294
313


Fab/CD3-scFv






DLL3-
3D43I
269
293
311


Fab/CD3-scFv






DLL3-
3D4B
262
291
295


Fab/CD3-scFv






DLL3-
3D4C
263
291
295


Fab/CD3-scFv






DLL3-
3D4D
264
291
295


Fab/CD3-scFv






DLL3-
3D4E
265
291
295


Fab/CD3-scFv






DLL3-
3D4F
266
291
295


Fab/CD3-scFv






DLL3-
3D4G
267
291
295


Fab/CD3-scFv






DLL3-
3D34-4C
341
291
295


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D35-4C
342
291
295


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D36-4C
346
291
295


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D37-4C
345
291
295


scFvxCD3-






scFv/DLL3-






Fab






DLL3-
3D4H
268
291
295


Fab/CD3-scFv






DLL3-
3D4I
269
291
295


Fab/CD3-scFv






DLL3-
3D4J
270
291
295


Fab/CD3-scFv






DLL3-
3D4K
271
291
295


Fab/CD3-scFv






DLL3-
3D4L
272
291
295


Fab/CD3-scFv






DLL3-
3D7
273
292
298


Fab/CD3-scFv






DLL3-
3D34-7C
341
292
298


scFvxCD3-






scFv/DLL3-






Fab
















TABLE 3







DLL3-CD28

















seq





Seq ID
Seq Id
id




Molecule
chain
chain
chain



format
Name
1
2
3







CD28-
28D1 
425
426
431



Fab/DLL3-







scFv







CD28-
28D2 
425
426
434



Fab/DLL3-







scFv







CD28-
28D3 
425
426
432



Fab/DLL3-







scFv







CD28-
28D4 
425
426
433



Fab/DLL3-







scFv







CD28-
28D9 
427
290
298



scFv/DLL3-







Fab







CD28-
28D10
427
291
295



scFv/DLL3-







Fab







CD28-
28D11
427
292
298



scFv/DLL3-







Fab







CD28-
28D12
427
292
296



scFv/DLL3-







Fab







CD28-
28D13
428
291
295



scFv/DLL3-







Fab







CD28-
28D14
429
291
295



scFv/DLL3-







Fab







CD28-
28D15
430
291
295



scFv/DLL3-







Fab







CD28-
28D16
430
292
296



scFv/DLL3-







Fab







CD28-
28D17
430
276
302



scFv/DLL3-







Fab







CD28-
28D18
430
290
298



scFv/DLL3-







Fab







CD28-
28D19
430
276
297



scFv/DLL3-







Fab

















TABLE 4







DLL4-41 BB












Molecule
Seq ID
seq ID
seq Id


format
Name
chain1
chain2
chain3





4-1BB-Fab/DLL3-scFv
4D1 
437
438
431


4-1BB-Fab/DLL3-scFv
4D2 
437
438
434


4-1BB-Fab/DLL3-scFv
4D3 
437
438
432


4-1BB-Fab/DLL3-scFv
4D4 
437
438
433


4-1BB-scFv/DLL3-Fab
4D5 
441
291
295


4-1BB-scFv/DLL3-Fab
4D6 
442
291
295


4-1BB-scFv/DLL3-Fab
4D7 
445
291
295


4-1BB-Fab/DLL3-scFv
4D8 
437
438
466


4-1BB-Fab/DLL3-scFv
4D9 
437
438
467


4-1BB-scFv/DLL3-Fab
4D10
446
291
295


4-1BB-scFv/DLL3-Fab
4D11
447
291
295


4-1BB-scFv/DLL3-Fab
4D12
448
291
295


4-1BB-scFv/DLL3-Fab
4D13
444
291
295


4-1BB-scFv/DLL3-Fab
4D14
443
290
298


DLL3-scFv/4-1BB-Fab
4D15
432
439
440


DLL3scFv-41BBscFv-fc 
4D16
468
439
440


X 41BBFab-fc






4-1BBscFv-41BBscFv-Fc
4D17
469
291
295


x DLL3Fab-Fe






41BBFab-Fe-DLL3scFv
4D18
470
472
440


41BBFab-Fe-DLL3scFv
4D19
471
473
298
















TABLE 5







DLL3 × CD3 × CD28












Molecule
Seq Id 
seq ID 
Seq ID 


format
Name
chain 1
Chain 2
chain 3





CD28scFvxCD3scFv/
328D1
481
276
302


DLL3-Fab






CD28scFvxCD3scFv/
328D2
481
291
295


DLL3-Fab






CD28scFvxCD3scFv/
328D3
481
290
298


DLL3-Fab






CD28scFvxCD3scFv/
328D4
481
276
297


DLL3-Fab






DLL3scFvxCD3scFv/
328D5
343
425
426


CD28-Fab




















TABLE 6







DLL3 × CD3 × CD137












Molecule
Seq ID 
Seq ID
Seq Id 


format
Name
chain 1
Chain2
chain 3





DLL3-scFvxCD3scFv/4-
34D1
343
437
438


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D2
341
437
438


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D3
342
437
438


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D4
343
437
438


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D5
346
439
440


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D6
341
439
440


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D7
342
439
440


1BB-Fab






DLL3-scFvxCD3scFv/4-
34D8
343
439
440


1BB-Fab






4-1BBFab-Fc-CD3scFv x
34D9
487
471
438


4-1BBFab-Fc-DLL3scFv






4-1BB-
 34D10
494
437
438


FabxFcxCD3scFvxDLL3-






scFv/4-1BB-Fab






4-1BB-
 34D11
495
472
440


FabxFcxCD3scFv/4-






1BB-FabxFcxDLL3-scFv






4-1BB-
 34D12
496
496
440


FabxFcxCD3scFvxDLL3-






scFv/4-1BB-Fab






CD3-scFvxFcx4-1BB-
 34D13
486
498
295


scFv/DLL3-Fab-Fc-4-






1BBscFv






CD3-scFv x DLL3Fab-Fc-
 34D14
269
499
295


4-1BBscFv-4-1BB-






scFv






DLL3scFv-CD3scFv-Fc-
 34D15
497
488
—


4-1BBscFv x Fc-4-






1BBscFv






DLL3-
 34D16
346
489
—


scFvxCD3scFv/Fcx4-






1BB-scFvx4-1BB-scFv
















TABLE 7







MUC17 VH and VL pairs
















VH 
VL 
HCDR1
HCDR2
HDCR3
LCDR1
LCDR2
LCDR3


AB Name
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
SeqID


















Muc17.7
146
149
92
106
111
118
121
124


Muc17.21
146
148
92
106
111
118
121
124


Muc17.22
146
150
92
106
111
118
121
124


Muc17.23
146
169
92
106
111
118
121
124


Muc17.24
146
147
92
106
111
118
121
126


Muc17.2
139
154
93
105
108
116
119
123


Muc17.1
127
166
94
103
108
116
119
122


Muc17.10
127
164
94
103
108
116
119
122


Muc17.11
127
162
94
103
108
116
119
122


muc17.12
127
165
94
103
108
116
119
122


Muc17.13
127
161
94
103
108
116
119
122


Muc17.14
127
160
94
103
108
116
119
122


Muc17.25
140
163
94
103
108
116
119
122


Muc17.26
140
160
94
103
108
116
119
122


Muc17.27
140
153
94
103
108
116
119
122


Muc17.28
140
155
94
103
108
116
119
122


Muc17.29
129
163
94
103
108
116
119
122


Muc17.30
129
160
94
103
108
116
119
127


Muc17.31
129
153
94
103
108
116
119
122


Muc17.8
127
153
94
107
108
116
119
122


Muc17.9
127
163
94
103
108
116
119
122


Muc17.3
141
156
95
104
112
117
120
123


Muc17.15
145
159
97
101
109
 115|
119
125


Muc17.16
145
152
97
101
109
115
119
125


Muc17.17
128
152
96
99
109
115
119
125


Muc17.18
145
151
97
101
109
114
119
125


Muc17.19
145
167
97
101
109
114
119
125


Muc17.31
136
168
98
101
109
115
119
125


Muc17.32
135
168
97
101
109
115
119
125


Muc17.33
135
159
97
101
109
115
119
125


Muc17.34
135
152
97
101
109
115
119
125


Muc17.35
133
168
97
101
109
115
119
125


Muc17.20
131
157
97
100
110
115
119
125


Muc17.36
134
168
97
101
109
115
119
125


Muc17.37
132
168
97
101
109
115
119
125


Muc17.38
137
168
97
101
109
115
119
125


Muc17.39
144
152
97
99
109
115
119
125


Muc17.4
145
168
97
101
109
115
119
125


Muc17.40
144
168
97
99
109
115
119
125


Muc17.41
144
167
97
99
109
114
119
125


Muc17.42
143
152
96
101
109
115
119
125


Muc17.43
143
168
96
101
109
115
119
125


Muc17.44
143
167
96
101
109
114
119
125


Muc17.45
130
159
97
101
109
115
119
125


Muc17.46
142
167
96
99
109
114
119
125


Muc17.47
142
168
96
99
109
115
119
125


Muc17.48
136
159
97
101
109
115
119
125


Muc17.5
131
159
97
100
110
115
119
125


Muc17.49
136
152
97
101
109
115
119
125
















TABLE 8







MUC17 VH × CD3















SEQ



Molecule
SEQID
SEQID
ID


format
Name
Chain1
Chain2
Chain3





Muc17-Fab/CD3-
3M23C
263
379
387


scFv






Muc17-Fab/CD3-
3M24C
263
379
388


scFv






Muc17-Fab/CD3-
3M25C
263
372
390


scFv






Muc17-Fab/CD3-
3M26C
263
372
389


scFv






Muc17-Fab/CD3-
3M27C
263
377
390


scFv






Muc17-Fab/CD3-
3M28C
263
371
390


scFv






Muc17-Fab/CD3-
3M29C
263
368
382


scFv






Muc17-scFvxCD3-
3M2C2
352




scFv-scfc






Muc17-Fab/CD3-
3M30C
263
368
383


scFv






Muc17-Fab/CD3-
3M31C
263
368
385


scFv






Muc17-Fab/CD3-
3M32C
263
368
384


scFv






Muc17-Fab/CD3-
3M33C
263
368
381


scFv






Muc17-Fab/CD3-
3M34C
263
370
394


scFv






Muc17-Fab/CD3-
3M35C
263
370
391


scFv






Muc17-Fab/CD3-
3M36C
263
370
387


scFv






Muc17-Fab/CD3-
3M37C
263
369
394


scFv






Muc17-Fab/CD3-
3M38C
263
369
391


scFv






Muc17-Fab/CD3-
3M39C
263
369
387


scFv






Muc17-Fab/CD3-
3M40C
263
379
394


scFv






Muc17-Fab/CD3-
3M41C
263
379
391


scFv






Muc17-Fab/CD3-
3M42C
263
379
387


scFv






Muc17-Fab/CD3-
3M43C
263
376
399


scFv






Muc17-Fab/CD3-
3M44C
263
376
386


scFv






Muc17-Fab/CD3-
3M45C
263
376
390


scFv






Muc17-Fab/CD3-
3M46C
263
380
399


scFv






Muc17-Fab/CD3-
3M47C
263
380
386


scFv






Muc17-Fab/CD3-
3M48C
263
380
390


scFv






Muc17-Fab/CD3-
3M49C
263
377
399


scFv






Muc17-Fab/CD3-
3M50C
263
377
386


scFv






Muc17-Fab/CD3-
3M51C
263
377
390


scFv






Muc17-Fab/CD3-
3M52C
263
369
397


scFv






Muc17-Fab/CD3-
3M53C
263
369
392


scFv






Muc17-Fab/CD3-
3M54C
263
369
396


scFv






Muc17-Fab/CD3-
3M55C
263
369
398


scFv






Muc17-scFv/CD3-
3M55D
264
369
400


Fab






Muc17-scFv/CD3-
3M55I
269
369
400


Fab






Muc17-scFv/CD3-
3M55K
271
369
400


Fab






Muc17-Fab/CD3-
3M56C
263
374
399


scFv






Muc17-Fab/CD3-
3M57C
263
375
399


scFv






Muc17-Fab/CD3-
3M58C
263
373
399


scFv






Muc17-Fab/CD3-
3M59C
263
378
399


scFv






Muc17-Fab/CD3-
3M60C
263
369
393


scFv






Muc17-scFvxCD3-
3M61C
350
239



scFv






Muc17-scFvxCD3-
3M62C
363
239



scFv






Muc17-scFvxCD3-
3M62D
362
239



scFv






Muc17-scFvxCD3-
3M62I
364
239



scFv






Muc17-scFvxCD3-
3M62K
365
239



scFv






Muc17-scFvxCD3-
3M63C
350
380
399


scFv/Muc17-Fab






Muc17-scFvxCD3-
3M64C
363
369
398


scFv/Muc17-Fab






Muc17-scFv/CD3-
3M65C
366
263
400


Fab






Muc17-scFv/CD3-
3M66C
367
263
400


Fab






Muc17-scFvxCD3-
3M67I
349
239



scFv






Muc17-scFvxCD3-
3M68I
354
239



scFv






Muc17-scFvxCD3-
3M8B7
351




scFv-scfc






1MU32scFv-

361
239



CD3scFv-Fe






1MU8AscFv-

353
239



CD3scFv-Fe






1MU32scFv(Y32F)-

360
239



CD3scFv-Fe






1MU32scFv(M34L)-

357
239



CD3scFv-Fe






1 MU32scFv(T58S)-

359
239



CD3scFv-Fe
















TABLE 9







MUC17 × CD28














Molecule
SeqID
SeqID
SeqID



format
Name
Chain1
Chain2
Chain3







Muc17-
28M1
427
368
384



Fab/CD28-







scFv







Muc17-
28M2
427
380
399



Fab/CD28-







scFv







Muc17-
28M3
427
369
398



Fab/CD28-







scFv







Muc17-
28M4
430
368
384



Fab/CD28-







scFv







Muc17-
28M5
430
380
399



Fab/CD28-







scFv







Muc17-
28M6
430
369
398



Fab/CD28-







scFv





















TABLE 10







MUC17 × 4188














Molecule
Seq ID
seq ID
seq Id



format
Name
chain1
chain2
chain3







Muc17-scfv/4-
4M1
366
437
438



1BB-







Fab







Muc17-
4M2
367
437
438



scfv/4-1BB-







Fab







Muc17-
4M3
474
437
438



scfv/4-1BB-







Fab







4-1BB-
4M4
451
369
398



scFv/Muc17-







Fab







4-1BB-
4M5
545
369
398



scFv/Muc17-







Fab







4-1BB-
4M6
450
369
398



scFv/Muc17-







Fab







4-1BB-
4M7
451
380
399



scFv/Muc17-







Fab







4-1BB-
4M8
452
380
399



scFv/Muc17-







Fab







4-1BB-
4M9
453
369
398



scFv/Muc17-







Fab







4-1BB-
 4M10
447
369
398



scFv/Muc17-







Fab







4-1BB-
 4M11
448
369
398



scFv/Muc17-







Fab







4-1BB-
 4M12
444
369
398



scFv/Muc17-







Fab







4-1BB-
 4M13
443
369
398



scFv/Muc17-







Fab







4-1BB-
 4M14
452
380
399



scFv/Muc17-







Fab







4-1BB-
 4M15
453
380
399



scFv/Muc17-







Fab

















TABLE 11







MUC17 × CD3 × CD28













Seq Id 
seq ID
Seq ID 


format
Molecule Name
chain 1
Chain 2
chain 3





CD3-
328M1
480
369
398


scFvxCD28-






scFv/Muc17-Fab






CD28-
328M2
481
369
398


scFvxCD3-






scFv/Muc17-Fab






CD28-
328M3
481
380
399


scFvxCD3-






scFv/Muc17-Fab






CD28-
328M4
481
368
384


scFvxCD3-






scFv/Muc17-Fab






Muc17-
328M5
362
482
483


scFvxCD3-






scFv/CD28-Fab




















TABLE 12







MUC17 × CD3 × CD137













Seq Id 
seq ID 
Seq ID 


format
Molecule Name
chain 1
Chain 2
chain 3





Muc17-
34M1
362
437
438


scFvxCD3-






scFv/4-1BB-Fab






4-1BB-
34M2
486
380
399


scFvxCD3-






scFv/Muc17-Fab






4-1BB-
34M3
484
380
399


scFvxCD3-






scFv/Muc17-Fab






Muc17-
34M4
362
439
440


scFvxCD3-






scFv/4-1BB-Fab
















TABLE 13







CLDN18.2 VH and HL pairs
















VH
VL
HCDR1
HCDR2
HDCR3
LCDR1
LCDR2
LCDR3


AB Name
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID


















CLDN182.1
203
208
175
180
186
188
191
195


CLDN182.2
202
207
174
179
187
188
191
194


CLDN182.3
198
209
170
181
185
188
191
196


CLDN182.7
198
207
170
181
185
188
191
196


cldn18.2.4
201
212
173
176
182
190
191
193


CLDN18.8
201
211
173
176
182
190
191
193


CLDN182.5
204
210
171
177
184
189
191
192


CLDN182.6
206
206
172
178
183
188
191
192


CLDN182.9
206
213
172
178
183
188
191
192


CLDN182.10
200
212
173
176
182
190
191
193


CLDN182.11
200
211
173
176
182
190
191
193


CLDN182.12
197
209
170
181
185
188
191
196


CLDN182.13
197
207
170
181
185
188
191
196


CLDN182.14
199
207
174
179
187
188
191
194


CLDN182.15
205
206
172
178
183
188
191
192


CLDN182.16
205
213
172
178
183
188
191
192
















TABLE 14







CLDN18.2 × CD3














Molecule
SEQID
SEQID
SEQID



format
Name
Chain 1
Chain 2
Chain 3







Cldn-
3C17C
263
407
414



Fab/CD3-







scFv







Cldn-
3C18C
263
405
418



Fab/CD3-







scFv







Cldn-
3C19C
263
405
417



Fab/CD3-







scFv







Cldn-
3C20C
263
404
418



Fab/CD3-







scFv







Cldn-
3C21C
263
404
417



Fab/CD3-







scFv







Cldn-
3C22C
263
402
415



Fab/CD3-







scFv







Cldn-
3C23C
263
402
411



Fab/CD3-







scFv







Cldn-
3C24C
263
401
415



Fab/CD3-







scFv







Cldn-
3C25C
263
401
411



Fab/CD3-







scFv







Cldn-
3C26C
263
408
416



Fab/CD3-







scFv







Cldn-
3C27C
263
406
413



F&W/CD3-







scFv







Cldn-
3C28C
263
403
413



F&W/CD3-







scFv







Cldn-
3C29C
263
410
412



F&W/CD3-







scFv







Cldn-
3C30C
263
410
419



F&W/CD3-







scFv







Cldn-
3C31C
263
409
412



F&W/CD3-







scFv







Cldn-
3C32C
263
409
419



F&W/CD3-







scFv







Cldn-
3C18D
264
405
418



F&W/CD3-







scFv







Cldn-
3C18I
269
405
418



F&W/CD3-







scFv







Cldn-
3C18K
271
405
418



F&W/CD3-







scFv







Cldn-
3C22D
264
402
415



F&W/CD3-







scFv







Cldn-
3C22I
269
402
415



F&W/CD3-







scFv







Cldn-
3C22K
271
402
415



F&W/CD3-







scFv







Cldn-
3C26D
264
408
416



F&W/CD3-







scFv







Cldn-
3C26I
269
408
416



F&W/CD3-







scFv







Cldn-
3C26K
271
408
416



F&W/CD3-







scFv







Cldn-
3C27D
264
406
413



F&W/CD3-







scFv







Cldn-
3C27I
269
406
413



F&W/CD3-







scFv







Cldn-
3C27K
271
406
413



Fab/CD3-







scFv







CLDN-
3CBM






scFvxCD3-







scFv-scfc

















TABLE 15







CLDN18.2 × CD28














Molecule
SeqID
SeqID
SeqID



format
Name
Chain1
Chain2
Chain3







Cldn-
28C1
427
405
418



Fab/CD28-







scFv







Cldn-
28C2
427
402
415



Fab/CD28-







scFv







Cldn-
28C3
427
408
416



Fab/CD28-







scFv







Cldn-
28C4
427
406
413



Fab/CD28-







scFv







Cldn-
28C5
430
405
418



Fab/CD28-







scFv







Cldn-
28C6
430
402
415



Fab/CD28-







scFv







Cldn-
28C7
430
408
416



Fab/CD28-







scFv







Cldn-
28C8
430
406
413



Fab/CD28-







scFv







Cldn-
28C9
435
408
416



Fab/CD28-







scFv







Cldn-
 28C10
436
408
416



Fab/CD28-







scFv

















TABLE 16







CLDN18.2 × 4188














Molecule
SeqID
SeqID
SeqID



format
Name
chain 1
chain 2
chain 3







Cldn-
4C1 
475
437
438



scfv/4-







1BB-Fab







Cldn-
4C2 
476
437
438



scfv/4-







1BB-Fab







Cldn-
4C3 
477
437
438



scfv/4-







1BB-Fab







Cldn-
4C4 
478
437
438



scfv/4-







1BB-Fab







Cldn-
4C5 
451
405
418



Fab/4-







1BB-scFv







Cldn-
4C6 
454
405
418



Fab/4-







1BB-scFv







Cldn-
4C7 
451
408
416



Fab/4-







1BB-scFv







Cldn-
4C8 
459
408
416



Fab/4-







1BB-scFv







Cldn-
4C9 
479
408
416



Fab/4-







1BB-scFv







Cldn-
4C10
464
408
416



Fab/4-







1BB-scFv







Cldn-
4C11
460
408
416



Fab/4-







1BB-scFv







Cldn-
4C12
463
408
416



Fab/4-







1BB-scFv







Cldn-
4C13
465
408
416



Fab/4-







1BB-scFv







Cldn-
4C14
461
408
416



Fab/4-







1BB-scFv







Cldn-
4C15
454
408
416



Fab/4-







1BB-scFv







Cldn-
4C16
457
408
416



Fab/4-







1BB-scFv







Cldn-
4C17
455
408
416



Fab/4-







1BB-scFv







Cldn-
4C18
458
408
416



Fab/4-







1BB-scFv







Cldn-
4C19
456
408
416



Fab/4-







1BB-scFv

















TABLE 17







CLDN18.2 × CD3 × CD28













Seq ID 
seq ID 
seq Id 


format
Molecule Name
chain1
chain2
chain3





CD3-
328C1
481
405
418


scFvxCD28-






scFv/Cldn-Fab






CD3-
328C2
481
402
415


scFvxCD28-






scFv/Cldn-Fab






CD3-
328C3
481
408
416


scFvxCD28-






scFv/Cldn-Fab






CD3-
328C4
481
406
413


scFvxCD28-






scFv/Cldn-Fab




















TABLE 18







CLDN18.2 × CD3 × CD137













Seq ID 
seq ID 
seq Id 


format
Molecule Name
chain1
chain2
chain3





CD3-scFvx4-
34C1
491
405
418


1BB-scFv/Cldn-






Fab






CD3-scFvx4-
34C2
490
405
418


1BB-scFv/Cldn-






Fab






4-1BB-
34C3
486
405
418


scFvxCD3-






scFv/Cldn-Fab






4-1BB-
34C4
484
405
418


scFvxCD3-






scFv/Cldn-Fab






Cldn-scFvxCD3-
34C5
492
437
438


scFv/4-1BB-Fab






Cldn-scFvxCD3-
34C6
493
437
438


scFv/4-1BB-Fab






Cldn-scFvxCD3-
34C7
492
439
440


scFv/4-1BB-Fab






Cldn-scFvxCD3-
34C8
493
439
440


scFv/4-1BB-Fab
















TABLE 19







CLDN18.2 × CD28 × CD137













Seq ID 
seq ID 
seq Id 


format
Molecule Name
chain1
chain2
chain3





CD28-scFvx4-
284C1
500
405
418


1BB-scFv/Cldn-






Fab






CD28-scFvx4-
284C2
501
405
418


1BB-scFv/Cldn-






Fab






4-1BB-
284C3
502
405
418


scFvxCD28-






scFv/Cldn-Fab






4-1BB-
284C4
503
405
418


scFvxCD28-






scFv/Cldn-Fab













EXAMPLES
The compositions and methods described herein will be further understood by reference to the following examples, which are intended to be purely exemplary. The compositions and methods described herein are not limited in scope by the exemplified embodiments, which are intended as illustrations of single aspects only. Any methods that are functionally equivalent are within the scope of the invention. Various modifications of the compositions and methods described herein in addition to those expressly described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications fall within the scope of the invention.
Example 1
Humanized Anti-DLL3 Antibodies Binding to Human and Cynomolgus DLL3 Expressing CHO Cells
To evaluate the ability of anti-DLL3 antibodies to bind, human serial dilutions of the anti-DLL3 antibodies were added to CHO-K1 cells (20,000 cells/well) over-expressing human or cyno DLL3. The mixtures were incubated at 4° C. for 20 minutes, washed 3 times, and stained with the secondary antibody, PE labeled F(ab′)2-Goat anti-human IgG Fe (Thermo H10104) at 4° C. for 20 minutes. Cells were washed and resuspended in 7-Amino-Actinomycin D (7-AAD) solution and fixed in 10% neutral buffered formalin solution for 15 minutes before analysis with the iQue Intellicyt system. FIG. 2A is a graph of the concentration (nM) versus the mean fluorescence intensity.
Four humanized antibodies were used in this example (DLL3.1, DLL3.2, DLL3.3, DLL3.4). The experiment was repeated with cynomolgus monkey DLL3 expressing CHO cells. FIG. 2B is a graph of the concentration (nM) of the antibodies versus the mean fluorescence intensity for binding to the cells.
Example 2
Anti-DLL3Scfv-Fc Binding to huDLL3 Expressing CHO Cells
The experiment was repeated to evaluate the ability of anti-DLL3scfv-Fc antibody fragment to bind human DLL3. FIG. 3 is a graph of the concentration (nM) of the DLL3 molecules versus the mean fluorescence intensity for binding to the cells. Six antibody fragments were used in this example (DLL3.1 VH-VL, DLL3.2 VH-VL, DLL3.3 VH-VL, DLL3.4 VH-VL, DLL3.1 VL-VH and DLL3.4 VL-VH).
Example 3
CD28Scfv-CD3scFv-Fc×DLL3-Fab-Fc Trispecific Molecule 328D3 Activates PBMCs to Secrete IFN Gamma in the Presence of CHO-DLL3 Cells to Higher Levels than the CD3Scfv-Fc×DLL3Fab-Fc Bispecific Molecule 3D11 or CD28Scfv-Fc×DLL3Fab-Fc Bispecific Molecule 28D18
FIG. 4 is a graph of concentration (nM) of the molecules versus the IFNgamma as detected by ELISA (absorbance at 650 nm). The ELISA utilized high protein-binding 96-well plates that were coated overnight with 50 uL of mouse anti-human IFNgamma capture antibody at a concentration of 2 μg/ml. Plates were washed with PBS+0.05% Tween-20 (PBST) and blocked with 200 ul PBS with 1% BSA. After washing, the coated plates were incubated for 120 minutes with 50 uL of the supernatant from the PBMC:Target cell cultures stimulated with the molecules. Plates were washed, and then incubated for 60 minutes with 50 μL of biotinylated mouse anti-human IFNgamma detection antibody at a concentration of 0.5 μg/ml. Plates were washed and incubated with streptavidin-HRP for 20 minutes. After washing, captured IFNgamma was quantified using 3,3′,5,5′-tetramethylbenzidine (TMB)
Example 4
CD28Scfv-CD3scFv-Fc×DLL3-Fab-Fc Trispecific Molecule 328D2 Activates PBMCs to Secrete IFNgamma in the Presence of CHO-DLL3 Cells to Higher Levels than the CD3Scfv-Fc×DLL3Fab-Fc Bispecific Molecule 3D41 or CD28Scfv-Fc×DLL3Fab-Fc Bispecific Molecule 28D15
FIG. 5 is a graph of concentration (nM) of the molecules versus the IFNg (absorbance at 650 nm) as detected by ELISA as described in Example 3.
Example 5
CD28Scfv-CD3scFv-Fc×DLL3-Fab-Fc Trispecific Molecule 328D1 Activates PBMCs to Secrete IFNgamma in the Presence of CHO-DLL3 Cells to Higher Levels than the CD3Scfv-Fc×DLL3Fab-Fc Bispecific Molecule 3D221 or CD28Scfv-Fc×DLL3Fab-Fc Bispecific Molecule 28D17.
FIG. 6 is a graph of concentration (nM) of the molecules versus the IFNg (absorbance at 650 nm) as detected by ELISA as described in Example 3.
Example 6
CD28Scfv-CD3scFv-Fc×DLL3-Fab-Fc Trispecific Molecules 328D4 and 328D5 Activate PBMCs to Secrete More IL-2 in the Presence of NCI-H82 Cells Greater than the CD3Scfv-Fc×DLL3Fab-Fc Bispecific, 3D39I
FIG. 7 is a graph of concentration (nM) versus the IFNg (absorbance at 650 nm). After 48 hours of culture, T cell activation was measured as an increase in IL-2 secreted into the medium. IL-2 secretion was measured using an IL-2 ELISA. The ELISA utilized high protein-binding 96-well plates that were coated overnight with 50 μL of mouse anti-human IL-2 capture antibody at a concentration of 2 μg/ml. Plates were washed with PBS+0.05% Tween-20 (PBST) and blocked with 200 μL PBS with 1% BSA. After washing, the coated plates were incubated for 120 minutes with 50 μL of the supernatant from the PBMC:Target cell cultures stimulated with bispecific molecules. Plates were washed, and then incubated for 60 minutes with 50 μL of biotinylated mouse anti-human IL-2 detection antibody at a concentration of 0.5 μg/ml. Plates were washed and incubated with streptavidin-HRP for 20 minutes. After washing, captured IL-2 was quantified using 3,3′,5,5′-tetramethylbenzidine (TMB).
Example 7
MUC17×CD3 Bispecifics Bind Muc17-CHOK1 and ASPC1 Cells
Seven humanized antibodies, 1MU11A, 1MU32A, 1MU36A, 1MU16A, 1MU37A, 1MU43A, and 1MU47A, were assessed for the ability to bind to CHO-K1 cells expressing the membrane proximal fragment of MUC17.
To evaluate the ability of anti-MUC17 antibodies to bind cell expressed MUC17, serial dilutions of the anti-MUC17 antibodies were added to the MUC17 expressing CHO-K1 cells at a concentration of 20,000 cells/well. The antibody:cell mixtures were incubated at 4° C. for 20 minutes, washed 3 times, and stained with the secondary antibody, PE labeled F(ab′)2-Goat anti-human IgG Fc (Thermo H10104) at 4° C. for 20 minutes. Cells were washed and resuspended in 7-Amino-Actinomycin D (7-AAD) solution and fixed in 10% neutral buffered formalin solution for 15 minutes before analysis with the iQue Intellicyt system. FIG. 11A is a graph of the concentration (nM) versus the mean fluorescence intensity (MFI) of binding to MUC17-CHO cells.
The experiment was repeated with APSC1 cells, which endogenously express full length human MUC17. FIG. 11B shows a graph of the concentration (nM) versus the mean fluorescence intensity (MFI) for the antibodies binding to the ASPC1 cells.
Example 8
Hu1MU11A and Hu1MU32A Retain Muc17 Binding Activity in the Scfv Format
FIG. 12 is a graph of concentration (nM) of the Fab-Fc and Scfv-Fc versions of the molecules versus absorbance at 650 nm for binding to MUC17.
Example 9
CD3×CD28×Muc17 Trispecific Molecules have Similar Maximum Killing of ASPC1 Cells by PBMCs as the CD3 Bispecific 3M62I
The experiment was conducted to evaluate the ability of human peripheral mononuclear (PBMCs) effector cells to kill Muc17 expressing CHO target cells when stimulated by MUC17×CD3×CD28 trispecific molecules. Cell cytotoxicity was determined as the amount of lactate dehydrogenase (LDH) released from damaged cells as a percent of the total LDH release with 1% Triton-X (Max Killing) added to the cell mixtures at time 0. For the cytotoxicity assays, target (MUC17-CHO or ASPC1) and effector cells (human PBMCs) were suspended in 200 ul of medium containing 10% serum at an effector to target ratio of 10:1, in a 96-well plate. Dilutions of the bispecific molecules were added to the cultures in triplicate. The 96-well plates were cultured at 37° C. for 48 hours. The cells were then centrifuged at 250×g for 10 min, and 100 μl of the supernatant plus were transferred into corresponding wells of an optically clear 96-well plate containing 100 μl LDH assay reagent per well. The plates were then incubated for up to 30 min at room temperature. The absorbance of all samples was measured at 490-500 nm using a microtiter plate reader. FIG. 13 is a graph of the concentration (nM) of the molecules versus % of maximum killing.
Example 10
Muc17×CD3×cD28 Trispecific Molecules 328M2, 328M 3, 328M 4, and 328M 5 Activate PBMCs in the Presence of ASPC1 Cells to Secrete IL-2 at Levels Higher than that of the CD3 Bispecific, 3M62I
FIG. 14 is a graph of concentration (nM) of the molecules versus absorbance at 650 nm from the ELISA to detect secreted IL-2 in the media as described in Example 6.
Example 11
CD3Scfv×CD28SScfv×MUC17-Fab trispecific molecules 328MI and 328M2 activate T-cells to kill MUC17-CHO cells better than the CD3 bispecific 3M55I, and similar to the combination of CD3 and CD28 bispecifics, 3M55I+28M3. FIG. 15 is a graph of the concentration (nM) of the molecules versus % of maximum killing.
Example 12
CD28Scfv×CD3Scfv×Muc17Fab Trispecific 328M2 Activates PBMCs to Express CD25 Only when in the Presence of Muc17 Expressing CHO Cells and Cannot Activate without Muc17 Target Cells. In Contrast the CD3scFv×CD28scFv×Muc17-Fab Trispecific 328M1 is Able to Non-Specifically Activate Cells in the Absence of Muc17, Similarly to the Bispecific Lacking Muc17 Binding, 328-1
FIG. 16A shows the expression of CD25(MFI) stimulated by increasing concentrations of the bispecific and trispecific molecules T cells when PBMCs are in the presence of MUC17 expressing CHO cells, showing both 328M1 and 328M2 active T cells to express CD25 similarly.
FIG. 16B shows the expression of CD25(MFI) stimulated by increasing concentrations of the bispecific and trispecific molecules T cells when PBMCs are in the presence of parental CHO cells, showing 328M1 stimulates T cells whereas 328M2 can not stimulate without MUC17 expressing cells. Similarly, 328-1, which lacks the anti-MUC17 Fab, also activates T cells in the absence of MUC17 target cells, whereas 328-2 cannot.
Versions-1 have the configuration CD3Scfv-CD28Scfv-Fc×MUC17Fab-Fc or Fc and versions-2 have the configuration CD28scfv-CD3Scfv-Fc×MUC17Fab-Fc or Fc. This indicates the version-2 configuration, CD28scfv-CD3Scfv-Fc, has improved properties over the version-1 configuration, CD3Scfv-CD28Scfv-Fc, because version-2 configuration requires the presence of target cells to activate T cells and thus is less likely to cause non-specific activation of T cells and potential toxicity.
Example 13
CD137×CD3×Muc17 Trispecific Molecules 34M1 and 34M4 Increase CD4 and CD8 T Cell Numbers when PBMCs are Cultured with Muc17 Expressing CHO Cells to a Greater Level than the Benchmark CD3 Bispecific 3M8B7
FIG. 17A is a graph of concentration of the molecules in nM (treatment (nM)) versus CD8+ cell counts as determined by staining the cells with fluorescently labeled antibodies against CD3 and CD4 or CD8 and assessment by flow cytometry.
FIG. 17B is a graph of concentration of the molecules in nM (treatment (nM)) versus CD4+ cell counts as determined by staining the cells with fluorescently labeled antibodies against CD3 and CD4 or CD8 and assessment by flow cytometry.
Example 14
Trispecific Molecules 328C1, 328C 2, 328C3, 328C4 Activate T-Cells to Secrete IL-2 Better to a Greater Level than the 3C27I CD3×CIdn18.2 Bite Alone.
FIG. 21 is a graph of concentration (nM) of the molecules versus concentration of secreted IL-2 (pg/mL) as determined by ELISA.
Example 15
CD3×CD137×CLDN18.2 Trispecific Molecules 34C4 and 34C3 Activate PBMCs in the Presence of SNU-601 Cells to Express More CD25 than 3C18I
FIG. 22 is a graph of the concentration (nM) of the molecules versus CD25 mean fluorescence intensity (MFI) on CD3+ T cells from the cultures.
Example 16
CD3×CD137×CLDN18.2 Trispecific Molecules 34C4 and 34C3 Activate PBMCs in the Presence of SNU-601 Cells to Secrete More IFNgamma than 3C18I Alone
FIG. 23 is a graph of the concentration (nM) of the molecules versus IFNg (absorbance at 650 nm) as determined by ELISA.
Example 17
CD3×CD137×CLDN18.2 Trispecific 34C3 Increases CD8 T Cell Numbers Beyond What is Stimulated by the CD3 T Cell Engager 3C27I in the Presence of CHO Cells Expressing CLDN18.2
FIG. 24 is a graph of the concentration of the molecules in nM (treatment (nM)) versus CD8+ cell counts as determined by flow cytometry from the cultures.
The therapeutic method of the present specification may include the step of administering the composition including the antibody at a pharmaceutically effective amount. The total daily dose should be determined through appropriate medical judgment by a physician, and administered once or several times. The specific therapeutically effective dose level for any particular patient may vary depending on various factors well known in the medical art, including the kind and degree of the response to be achieved, concrete compositions according to whether other agents are used therewith or not, the patient's age, body weight, health condition, gender, and diet, the time and route of administration, the secretion rate of the composition, the time period of therapy, other drugs used in combination or coincident with the composition disclosed herein, and like factors well known in the medical arts.
In still another aspect, the present specification provides a use of the therapeutic protein or the pharmaceutical composition including the same in the preparation of drugs for the prevention or treatment of cancer, a neurodegenerative or an infectious disease.
In one embodiment, the dose of the composition may be administered daily, semi-weekly, weekly, bi-weekly, or monthly. The period of treatment may be for a week, two weeks, a month, two months, four months, six months, eight months, a year, or longer. The initial dose may be larger than a sustaining dose. In one embodiment, the dose ranges from a weekly dose of at least 0.01 mg/kg, at least 0.25 mg/kg, at least 0.3 mg/kg, at least 0.5 mg/kg, at least 0.75 mg/kg, at least 1 mg/kg, at least 2 mg/kg, at least 3 mg/kg, at least 4 mg/kg, at least 5 mg/kg, at least 6 mg/kg, at least 7 mg/kg, at least 8 mg/kg, at least 9 mg/kg, at least 10 mg/kg, at least 15 mg/kg, at least 20 mg/kg, at least 25 mg/kg, or at least 30 mg/kg In one embodiment, a weekly dose may be at most 1.5 mg/kg, at most 2 mg/kg, at most 2.5 mg/kg, at most 3 mg/kg, at most 4 mg/kg, at most 5 mg/kg, at most 6 mg/kg, at most 7 mg/kg, at most 8 mg/kg, at most 9 mg/kg, at most 10 mg/kg, at most 15 mg/kg, at most 20 mg/kg, at most 25 mg/kg, or at most 30 mg/kg. In a particular aspect, the weekly dose may range from 5 mg/kg to 20 mg/kg. In an alternative aspect, the weekly dose may range from 10 mg/kg to 15 mg/kg.
The present specification also provides a pharmaceutical composition for the administration to a subject. The pharmaceutical composition disclosed herein may further include a pharmaceutically acceptable carrier, excipient, or diluent. As used herein, the term “pharmaceutically acceptable” means that the composition is sufficient to achieve the therapeutic effects without deleterious side effects, and may be readily determined depending on the type of the diseases, the patient's age, body weight, health conditions, gender, and drug sensitivity, administration route, administration mode, administration frequency, duration of treatment, drugs used in combination or coincident with the composition disclosed herein, and other factors known in medicine.
The pharmaceutical composition including the antibody disclosed herein may further include a pharmaceutically acceptable carrier. For oral administration, the carrier may include, but is not limited to, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersing agent, a stabilizer, a suspending agent, a colorant, and a flavorant. For injectable preparations, the carrier may include a buffering agent, a preserving agent, an analgesic, a solubilizer, an isotonic agent, and a stabilizer. For preparations for topical administration, the carrier may include a base, an excipient, a lubricant, and a preserving agent.
The disclosed compositions may be formulated into a variety of dosage forms in combination with the aforementioned pharmaceutically acceptable carriers. For example, for oral administration, the pharmaceutical composition may be formulated into tablets, troches, capsules, elixirs, suspensions, syrups or wafers. For injectable preparations, the pharmaceutical composition may be formulated into an ampule as a single dosage form or a multidose container. The pharmaceutical composition may also be formulated into solutions, suspensions, tablets, pills, capsules and long-acting preparations.
On the other hand, examples of the carrier, the excipient, and the diluent suitable for the pharmaceutical formulations include, without limitation, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oils. In addition, the pharmaceutical formulations may further include fillers, anti-coagulating agents, lubricants, humectants, flavorants, and antiseptics.
Further, the pharmaceutical composition disclosed herein may have any formulation selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, liquids for internal use, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, lyophilized formulations and suppositories.
The composition may be formulated into a single dosage form suitable for the patient's body, and preferably is formulated into a preparation useful for peptide drugs according to the typical method in the pharmaceutical field so as to be administered by an oral or parenteral route such as through skin, intravenous, intramuscular, intra-arterial, intramedullary, intramedullary, intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal, intranasal, intracolonic, topical, sublingual, vaginal, or rectal administration, but is not limited thereto.
The composition may be used by blending with a variety of pharmaceutically acceptable carriers such as physiological saline or organic solvents. In order to increase the stability or absorptivity, carbohydrates such as glucose, sucrose or dextrans, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers may be used.
The administration dose and frequency of the pharmaceutical composition disclosed herein are determined by the type of active ingredient, together with various factors such as the disease to be treated, administration route, patient's age, gender, and body weight, and disease severity.
The total effective dose of the compositions disclosed herein may be administered to a patient in a single dose, or may be administered for a long period of time in multiple doses according to a fractionated treatment protocol. In the pharmaceutical composition disclosed herein, the content of active ingredient may vary depending on the disease severity. Preferably, the total daily dose of the peptide disclosed herein may be approximately 0.0001 μg to 500 mg per 1 kg of body weight of a patient. However, the effective dose of the peptide is determined considering various factors including patient's age, body weight, health conditions, gender, disease severity, diet, and secretion rate, in addition to administration route and treatment frequency of the pharmaceutical composition. In view of this, those skilled in the art may easily determine an effective dose suitable for the particular use of the pharmaceutical composition disclosed herein. The pharmaceutical composition disclosed herein is not particularly limited to the formulation, and administration route and mode, as long as it shows suitable effects.
Moreover, the pharmaceutical composition may be administered alone or in combination or coincident with other pharmaceutical formulations showing prophylactic or therapeutic efficacy.
In still another aspect, the present specification provides a method for preventing or treating of cancer, infectious diseases or neurodegenerative diseases comprising the step of administering to a subject the chimeric protein or the pharmaceutical composition including the same.
Given the teachings and guidance provided herein, those skilled in the art will understand that a formulation described herein can be equally applicable to many types of biopharmaceuticals, including those exemplified, as well as others known in the art. Given the teachings and guidance provided herein, those skilled in the art also will understand that the selection of, for example, type(s) or and/or amount(s) of one or more excipients, surfactants and/or optional components can be made based on the chemical and functional compatibility with the biopharmaceutical to be formulated and/or the mode of administration as well as other chemical, functional, physiological and/or medical factors well known in the art. For example, non-reducing sugars exhibit favorable excipient properties when used with polypeptide biopharmaceuticals compared to reducing sugars. Accordingly, exemplary formulations are exemplified further herein with reference to polypeptide biopharmaceuticals. However, the range of applicability, chemical and physical properties, considerations and methodology applied to polypeptide biopharmaceutical can be similarly applicable to biopharmaceuticals other than polypeptide biopharmaceuticals.
In various embodiments, a formulation can include, without limitation, combinations of bioactive agents (such as viruses, proteins, antibodies, peptides and the like as described herein) in the formulation. For example, a formulation as described herein can include a single bioactive agent for treatment of one or more conditions, including without limitation, disease. A formulation as described herein also can include, in an embodiment, without limitation, two or more different bioactive agents for a single or multiple conditions. Use of multiple bioactive agents in a formulation can be directed to, for example, the same or different indications. Similarly, in another embodiment, multiple bioactive agents can be used in a formulation to treat, for example, both a pathological condition and one or more side effects caused by the primary treatment. In a further embodiment, multiple bioactive agents also can be included, without limitation, in a formulation as described herein to accomplish different medical purposes including, for example, simultaneous treatment and monitoring of the progression of the pathological condition. In an additional embodiment, multiple, concurrent therapies such as those exemplified herein as well as other combinations well known in the art are particularly useful for patient compliance because a single formulation can be sufficient for some or all suggested treatments and/or diagnosis. Those skilled in the art will know those bioactive agents that can be admixed for a wide range of combination therapies. Similarly, in various embodiments, a formulation can be used with a small molecule drug and combinations of one or more bioactive agents together with one or more small molecule pharmaceuticals. Therefore, in various embodiments a formulation is provided containing 1, 2, 3, 4, 5 or 6 or more different bioactive agents, as well as, for one or more bioactive agents combined with one or more small molecule pharmaceuticals.
In various embodiments, a formulation can include, one or more preservatives and/or additives known in the art. Similarly, a formulation can further be formulated, without limitation, into any of various known delivery formulations. For example, in an embodiment, a formulation can include, surfactants, adjuvant, biodegradable polymers, hydrogels, etc., such optional components, their chemical and functional characteristics are known in the art. Similarly known in the art are formulations that facilitate rapid, sustained or delayed release of the bioactive agents after administration. A formulation as described can be produced to include these or other formulation components known in the art.
The composition can therefore be administered as a single dose, or as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via an implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. Appropriate dosages may be ascertained through use of appropriate dose-response data. In various embodiments, the bioactive agents in formulations described herein can, without limitation, be administered to patients throughout an extended time period, such as chronic administration for a chronic condition. The composition can be a solid, a semi-solid or an aerosol and a pharmaceutical compositions is formulated as a tablet, geltab, lozenge, orally dissolved strip, capsule, syrup, oral suspension, emulsion, granule, sprinkle or pellet.
In an embodiment, for oral, rectal, vaginal, parenteral, pulmonary, sublingual and/or intranasal delivery formulations, tablets can be made by compression or molding, optionally with one or more accessory ingredients or additives. In an embodiment, compressed tablets are prepared, for example, by compressing in a suitable tabletting machine, the active ingredients in a free-flowing form such as a powder or granules, optionally mixed with a binder (for example, without limitation, povidone, gelatin, hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, without limitation, sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) and/or surface-active or dispersing agent.
In an embodiment, molded tablets are made, for example, without limitation, by molding in a suitable tabletting machine, a mixture of powdered compounds moistened with an inert liquid diluent. In an embodiment, the tablets may optionally be coated or scored, and may be formulated so as to provide slow or controlled release of the active ingredients, using, for example, without limitation, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. In an embodiment, tablets may optionally be provided with a coating, without limitation, such as a thin film, sugar coating, or an enteric coating to provide release in parts of the gut other than the stomach. In an embodiment, processes, equipment, and toll manufacturers for tablet and capsule making are well-known in the art.
In an embodiment, capsule formulations can utilize either hard or soft capsules, including, without limitation, gelatin capsules or vegetarian capsules such as those made out of hydroxymethylpropylcellulose (HMPC). In an embodiment, a type of capsule is a gelatin capsule. In an embodiment, capsules may be filled using a capsule filling machine such as, without limitation, those available from commercial suppliers such as Miranda International or employing capsule manufacturing techniques well-known in the industry, as described in detail in Pharmaceutical Capsules, 2.sup.nd Ed., F. Podczeck and B. Jones, 2004. In an embodiment, capsule formulations may be prepared, without limitation, using a toll manufacturing center such as the Chao Center for Industrial Pharmacy & Contract Manufacturing, located at Purdue Research Park.
Packaging and instruments for administration may be determined by a variety of considerations, such as, without limitation, the volume of material to be administered, the conditions for storage, whether skilled healthcare practitioners will administer or patient self-compliance, the dosage regime, the geopolitical environment (e.g., exposure to extreme conditions of temperature for developing nations), and other practical considerations.
Injection devices include pen injectors, auto injectors, safety syringes, injection pumps, infusion pumps, glass prefilled syringes, plastic prefilled syringes and needle free injectors syringes may be prefilled with liquid, or may be dual chambered, for example, for use with lyophilized material. An example of a syringe for such use is the Lyo-Ject™, a dual-chamber pre-filled lyosyringe available from Vetter GmbH, Ravensburg, Germany. Another example is the LyoTip which is a prefilled syringe designed to conveniently deliver lyophilized formulations available from LyoTip, Inc., Camarillo, California, U.S.A. Administration by injection may be, without limitation intravenous, intramuscular, intraperitoneal, or subcutaneous, as appropriate. Administrations by non-injection route may be, without limitation, nasal, oral, cocular, dermal, or pulmonary, as appropriate.
In certain embodiments, kits can comprise, without limitation, one or more single or multi-chambered syringes (e.g., liquid syringes and lyosyringes) for administering one or more formulations described herein. In various embodiments, the kit can comprise formulation components for parenteral, subcutaneous, intramuscular or IV administration, sealed in a vial under partial vacuum in a form ready for loading into a syringe and administration to a subject. In this regard, the composition can be disposed therein under partial vacuum. In all of these embodiments and others, the kits can contain one or more vials in accordance with any of the foregoing, wherein each vial contains a single unit dose for administration to a subject.
The kits can comprise lyophilates, disposed as herein, that upon reconstitution provide compositions in accordance therewith. In various embodiment the kits can contain a lyophilate and a sterile diluent for reconstituting the lyophilate.
Also described herein, are methods for treating a subject in need of therapy, comprising administering to the subject an effective amount of a formulation as described herein. The therapeutically effective amount or dose of a formulation will depend on the disease or condition of the subject and actual clinical setting.
In an embodiment, a formulation as described herein can be administered by any suitable route, specifically by parental (including subcutaneous, intramuscular, intravenous and intradermal) administration. It will also be appreciated that the preferred route will vary with the condition and age of the recipient, and the disease being treated. Methods of determining the most effective means and dosage of administration are known to those of skill in the art and will vary, without limitation, with the composition used for therapy, the purpose of the therapy, and the subject being treated. Single or multiple administrations can be carried out, without limitation, the dose level and pattern being selected by the treating physician. Suitable dosage formulations and methods of administering the agents are known in the art.
The formulations as described herein can be used in the manufacture of medicaments and for the treatment of humans and other animals by administration in accordance with conventional procedures.
Also provided herein are combinatorial methods for developing suitable virus formulations using combinations of amino acids. These methods are effective for developing stable liquid or lyophilized formulations, and particularly pharmaceutical virus formulations.
Compositions in accordance with embodiments described herein have desirable properties, such as desirable solubility, viscosity, syringeability and stability. Lyophilates in accordance with embodiments described herein have desirable properties, as well, such as desirable recovery, stability and reconstitution.
In an embodiment, the pH of the pharmaceutical formulation is at least about 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, or 9.
In an embodiment, the pH of the pharmaceutical formulation is from about 3 to about 9, about 4 to about 19, about 5 to about 9, about 6 to about 8, about 6 to about 7, about 6 to about 9, about 5 to about 6, about 5 to about 7, about 5 to about 8, about 4 to about 9, about 4 to about 8, about 4 to about 7, about 4 to about 6, about 4 to about 5, about 3 to about 8, about 3 to about 7, about 3 to about 6, about 3 to about 5, about 3 to about 4, about 7 to about 8, about 7 to about 9, about 7 to about 10.
Certain embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the present invention to be practiced otherwise than specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described embodiments in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.
Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.
Unless otherwise indicated, all numbers expressing a characteristic, item, quantity, parameter, property, term, and so forth used in the present specification and claims are to be understood as being modified in all instances by the term “about.” As used herein, the term “about” means that the characteristic, item, quantity, parameter, property, or term so qualified encompasses a range of plus or minus ten percent above and below the value of the stated characteristic, item, quantity, parameter, property, or term. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical indication should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and values setting forth the broad scope of the invention are approximations, the numerical ranges and values set forth in the specific examples are reported as precisely as possible. Any numerical range or value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Recitation of numerical ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate numerical value falling within the range. Unless otherwise indicated herein, each individual value of a numerical range is incorporated into the present specification as if it were individually recited herein.
The terms “a,” “an,” “the” and similar referents used in the context of describing the present invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the present invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the present specification should be construed as indicating any non-claimed element essential to the practice of the invention.
Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term “consisting of” excludes any element, step, or ingredient not specified in the claims. The transition term “consisting essentially of” limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s). Embodiments of the present invention so claimed are inherently or expressly described and enabled herein.
Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.
All patents, patent publications, and other publications referenced and identified in the present specification are individually and expressly incorporated herein by reference in their entirety for the purpose of describing and disclosing, for example, the compositions and methodologies described in such publications that might be used in connection with the present invention. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.
In closing, it is to be understood that although aspects of the present specification are highlighted by referring to specific embodiments, one skilled in the art will readily appreciate that these disclosed embodiments are only illustrative of the principles of the subject matter disclosed herein. Therefore, it should be understood that the disclosed subject matter is in no way limited to a particular methodology, protocol, and/or reagent, etc., described herein. As such, various modifications or changes to or alternative configurations of the disclosed subject matter can be made in accordance with the teachings herein without departing from the spirit of the present specification. Lastly, the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. Accordingly, the present invention is not limited to that precisely as shown and described.







TABLE 20







SEQUENCES









Seq ID
sequence
name












1
DYIFSNYYIE
HCDR1;




DLL3.2,




DLL3.4


 


2
DYTFSNYYIE
HCDR1;




DLL3.1


 


3
DYYMN
HCDR1,




DLL3.9


 


4
DYYVN
HCDR1;




DLL3.8


 


5
GFTFSNYGMH
HCDR1:




DLL3.10,




DLL3.27,




DLL3.29,




DLL3.30,




DLL3.31,




DLL3.32,




DLL3.33,




DLL3.38


 


6
GFTFSSYGMH
HCDR1:




DLL3.5,




DLL3.12




DLL3.13,




DLL3.14,




DLL3.15,




DLL3.16,




DLL3.17




DLL3.18,




DLL3.19.




DLL3.20,




DLL3.21,




DLL3.22,




DLL3.23,




DLL3.24,




DLL3.25,




DLL3.26


 


7
GFTFSSYGMH
HCDR1:




DLL3.3,




DLL3.11


 


8
SAYYWN
HCDR1:




DLL23.34,




DLL3.35


 


9
SYYWS
HCDR1:




DDL3.28


 


10
EILPGNGNTVYNEKFKD
HCDR2;




DLL3.1


 


11
EILPGTGNTVYNEKFKD
HCDR2:




DLL3.2,




DLL3.4


 


12
IISPNDGGTNYNQKFKG
HCDR2;




DLL3.8


 


13
VINPDNGITTYNQKFKG
HCDR2;




DLL3.9


 


14
VINPYNDITIYNQKFQG
HCDR2:




DLL3.3


 


15
VISGSGSSKYYADSVKG
HCDR2;




DLL3.11,




DLL3.15,




DLL3.18,




DLL3.22


 


16
VISHHGSSKYYADSVKG
HCDR2:




DLL3.5,




DLL3.10,




DLL3.12,




DLL3.14,




DLL3.17,




DLL3.19,




DLL3.21,




DLL3.23,




DLL3.25,




DLL3.26,




DLL3.27,




DLL3.31,




DLL3.32,




DLL3.33


 


17
VISHHGSSKYYARSVKG
HCDR2:




DLL3.29,




DLL3.30,




DLL3.36


 


18
VISYDGSSKYYADSVKG
HCDR2;




DLL3.13,




DLL3.16,




DLL3.20,




DLL3.24


 


19
YISDVGHNYYNPSLKN
HCDR2;




DLL3.34


 


20
YISDVGSNNYNPSLKN
HCDR2;




DLL3.35


 


21
YVYYSGTTNYNPSLKS
HCDR2;




DLL3.28


 


22
WGDYALFAN
HCDR3:




DLL3.1,




DLL3.2,




DLL3.4


 


23
DQVFAY
HCDR3;




DLL3.8,




DLL3.34,




DLL3.35


 


24
DWFFYLFDY
HCDR3;




DLL3.5,




DLL3.10,




DLL3.11,




DLL3.12,




DLL3.13,




DLL3.14,




DLL3.15,




DLL3.16,




DLL3.17,




DLL3.26,




DLL3.27,




DLL3.31


 


25
DWFYFIFDY
HCDR3;




DLL3.18,




DLL3.19,




DLL3.20,




DLL3.21,




DLL3.22,




DLL3.23,




DLL3.24,




DLL3.25,




DLL3.32,




DLL3.33


 


26
DWWELVFDY
HCDR3;




DLL3.29,




DLL3.20,




DLL3.36


 


27
EGVLYDGYYEGAY
HCDR3;




DLL3.3


 


28
GVWNYERSFDY
HCDR3;




DLL3.9


 


29
SIAVTGFYFDY
HCDR3;




DLL3.28


 


30
SASSSVSYMH
LCDR1;




DLL3.35


 


31
KASQNVGIAVA
LCDR1;




DLL3.3


 


32
KASQNVGTNVA
LCDR1;




DLL3.1,




DLL3.2,




DLL3.4,




DLL3.9,




DLL3.28


 


33
KSSQSLLHSDAKTFLY
LCDR1;




DLL3.22,




DLL3.23,




DLL3.24,




DLL3.25,




DLL3.26,




DLL3.27,




DLL3.32,




DLL3.33


 


34
KSSQSLLHSDGKTFLY
LDCR1:




DLL3.5,




DLL3.10,




DLL3.15,




DLL3.16,




DLL3.17,




DLL3.29,




DLL3.30,




DLL3.32,




DLL3.36


 


35
RASESVHSYGNSLIH
LDCR1;




DLL3.34


 


36
RSSKSLLHSNGITYLY
LCDR1;




DLL3.8


 


37
RSSOSLLHSDAKTFLD
LCDR1;




DLL3.18,




DLL3.19,




DLL3.20,




DLL3.21,


 


38
RSSQSLLHSDGKTFLD
LCDR1:




DLL3.11,




DLL3.12,




DLL3.13,




DLL3.14


 


39
SASYRYS
LCDR2:




DLL3.4,




DLL3.28


 


40
AASNRYT
LCDR2:




DLL3.3


 


41
DTSKLAS
LCDR2;




DLL3.35


 


42
EVSNRAS
LCDR2;




DLL3.11,




DLL3.12,




DLL3.13,




DLL3.14,




DLL3.18,




DLL3.19,




DLL3.20,




DLL3.21


 


43
EVSNRFS
LCDR2:




DLL3.5,




DLL3.10,




DLL3.15,




DLL3.16,




DLL3.17,




DLL3.22,




DLL3.23,




DLL3.14,




DLL3.25,




DLL3.26,




DLL3.27,




DLL3.29,




DLL3.30,




DLL3.31,




DLL3.32,




DLL3.33,




DLL3.36


 


44
QMSNLAS
LCDR2;




DLL3.8


 


45
RASNLES
LCDR2;




DLL3.34


 


46
SASYRYS
LCDR2;




DLL3.1,




DLL3.2,




DLL3.9


 


47
QQYSTYPYT
LCDR3:




DLL3.3


 


48
AQNLELP
LCDR3:




DLL3.8


 


49
LQGERLPFT
LCDR3;




DLL3.5,




DLL3.10,




DLL3.11,




DLL3.12,




DLL3.13,




DLL3.14,




DLL3.15,




DLL3.16,




DLL3.17,




DLL3.26,




DLL3.27,




DLL3.31


 


50
LQGIHLPFT
LCDR3;




DLL3.29,




DLL3.30,




DLL3.36


 


51
LQGRELPFT
LCDR3;




DLL3.18,




DLL3.19,




DLL3.20,




DLL3.21,




DLL3.22,




DLL3.23,




DLL3.24,




DLL3.25,




DLL3.32,




DLL3.33


 


52
QQTNEDP
LCDR3;




DLL3.34


 


53
QQWSSNPLT
LCDR3;




DLL3.35


 


54
QQYNNYPLT
LCDR3;




DLL3.4,




DLL3.9,




DLL3.28


 


55
QQYNSYPFT
LCDR3;




DLL3.1,




DLL3.2


 


56
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH;



GLEWVAVISGSGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.11,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSS
DLL3.15


 


57
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH:



GLEWVAVISGSGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.18,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSS
DLL3.22


 


58
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH; DLL3.5,



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.12,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSS
DLL3.26


 


59
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH;



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.19,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSS
DLL3.23


 


60
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH;



GLEWVAVISYDGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.13,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSS
DLL3.16,


 


61
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH;



GLEWVAVISYDGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.20,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSS
DLL3.24


 


62
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH;



GLEWVSVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.14,



EDTAVYYCAKDWFFYLFDYWGQGTLVTVSS
DLL3.17


 


63
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
VH;



GLEWVSVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.21,



EDTAVYYCAKDWFYFIFDYWGQGTLVTVSS
DLL3.25


 


64
EVQLQQSGPVLVKPGASVKMSCKASGFTFTDYYMNWVKQSHGK
VH; DLL3.9



SLEWIGVINPDNGITTYNQKFKGKATLTVDKSSSTAYMELNGLTSE




DSAVYYCARGVWNYERSFDYWGQGTTLTVSS



 


65
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGK
VH



GLEWVAVISHHGSSKYYADSVKGRYTISRDNSKNTLYLQMNSLRA
DLL3.27



EDTAVYYCARDWFFYLFDYWGQGTLVTVSS



 


66
QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGL
VH;



EWIGYVYYSGTTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTA
DLL3.28



VYYCASIAVTGFYFDYWGQGTLVTVSS



 


67
QVQLQQSGPVLVKPGASVKMSCKASGYSFTDYYVNWVKQSHGK
VH; DLL3.8



SLEWIGIISPNDGGTNYNQKFKGKATLTVDKSSSTAYMEVNSLTSE




DSAVYYCARDDDLGWYFDVWGTGTTVTVSS



 


68
QVQLVESGGGAVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
VH



GLEWVAVISHHGSSKYYARSVKGRFTISRDNSKNTLYLEMNSLRA
DLL3.29,



EDTAVYYCARDWWELVFDYWGQGTLVTVSS
DLL3.30,




DLL3.36


 


69
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
VH



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.10,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSS
DLL3.31


 


70
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
VH;



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.32,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSS
DLL3.33


 


71
QVQLVQSGAEVKKPGASVKVSCKASDYTFSNYYIEWVRQAPGQG
VH; DLL3.1



LEWMGEILPGNGNTVYNEKFKDRVTMTVDTSTSTAYMELRSLRSD




DTAVYYCARWGDYALFANWGQGTLVTVSS



 


72
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
VH; DLL3.3



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS




EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSS



 


73
QVQLVQSGAEVKKPGASVKVSCKATDYIFSNYYIEWVRQAPGQGL
VH; DLL3.2,



EWMGEILPGTGNTVYNEKFKDRVTMTVDTSTSTVYMELSSLRSED
DLL3.4



TAVYYCARWGDYALFANWGQGTLVTVSS



 


74
SDVQLQESGPGLVKPSQSLSLTCSVTGYSITSAYYWNWIRQFPGN
VH DLL3.34



KLEWMGYISDVGHNYYNPSLKNRISITRDTSKNQFFLKLNSVTPED




TATYYCARDQVFAYWGQGTLVTVSA



 


75
SDVQLQESGPGLVKPSQSLSLTCSVTGYSITSAYYWNWIRQFPGN
VH DLL3.35



KLEWMGYISDVGSNNYNPSLKNRISITRDTFKNQFFLKLNSVTTED




TATYFCTRDQVFAYWGQGTLVTVS



 


76
DIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQKPGKAPKL
VL; DLL3.3



LIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYS




TYPYTFGQGTKLEIK



 


77
DIQMTQSPSFLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPK
VL; DLL3.4



PLIYSTSYRYSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQY




NNYPLTFGGGTKVEIK



 


78
DIQMTQSPSSLSASVGDRVTITCKSSQSLLHSDAKTFLYWYQQKP
VL; DLL3.26



GKAPKLLIYEVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATY




YCLQGERLPFTFGQGTKVEIK



 


78
DIQMTQSPSSLSASVGDRVTITCKSSQSLLHSDAKTFLYWYQQKP
VL; DLL3.27



GKAPKLLIYEVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATY




YCLQGERLPFTFGQGTKVEIK



 


79
DIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPK
VL; DLL3.1



ALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQY




NSYPFTFGQGTKLEIK



 


79
DIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPK
VL; DLL3.2



ALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQY




NSYPFTFGQGTKLEIK



 


80
DIVMTQAAFSNPVTVGTSASISCRSSKSLLHSNGITYLYWYLQKPG
VL; DLL3.8



QSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVY




YCAQNLELPWTFGGGTKLEIK



 


81
DIVMTQSPKFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSP
VL; DLL3.9



KALIYSASYRYSGVPDRFTGSGSGTDFTLTFSSVQSEDLAEYFCQ




QYNNYPLTFGGGTKLEIK



 


82
DIVMTQSPLSLPVTPGEPASISCKSSQSLLHSDAKTFLYWYLQKPG
VL; DLL3.22,



QSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.23,



CLQGRELPFTFGQGTKVEIK
DLL3.24,




DLL3.25


 


83
DIVMTQSPLSLPVTPGEPASISCKSSQSLLHSDGKTFLYWYLQKPG
VL; DLL3.5,



QSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.15,



CLQGERLPFTFGQGTKVEIK
DLL3.16,




DLL3.17


 


84
DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSDAKTFLDWYLQKPG
VL DLL3.18,



QSPQLLIYEVSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.19,



CLQGRELPFTFGQGTKVEIK
DLL3.20,




DLL3.21


 


85
DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSDGKTFLDWYLQKPG
VL;



QSPQLLIYEVSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.11,



CLQGERLPFTFGQGTKVEIK
DLL3.12,




DLL3.13,




DLL3.14


 


86
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDAKTFLYWYLQKPG
VL DLL3.32,



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.33



CLQGRELPFTFGPGTKVEIK



 


87
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPG
VL;



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.10,



CLQGERLPFTFGPGTKVEIK
DLL3.31


 


88
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPG
VL;



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.29,



CLQGIHLPFTFGPGTKVEIK
DLL3.30,




DLL3.36


 


89
DTVLTQSPASLAVSLGQRATISCRASESVHSYGNSLIHWYQQKPG
VL; DLL3.34



QPPRLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYC




QQTNEDPLTFGAGTKLELK



 


90
EIVLTQSPGTLSLSPGERVTLSCRASQRVNNNYLAWYQQRPGQAP
VL; DLL3.28



RLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQ




YDRSPLTFGGGTKLEIK



 


91
QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKR
VL; DLL3.35



WIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQW




SSNPLTFGAGTKLELK



 


92
GFTFSSFGMH
HCDR1;




Muc17.7,




Muc17.21,




Muc17.22,




Muc17.23,




Muc17.24


 


93
GYAFSDYWIN
HCDR1:




Muc17.2


 


94
GYEFSSHWMN
HCDR1;




Muc17.1,




Muc17.8,




Muc17.9,




muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14,




Muc17.25,




Muc71.26,




Muc17.27,




Muc17.28,




Muc17.29,




Muc17.30,




Muc17.31


 


95
GYIFSNHWMN
HCDR1:




Mcu17.3


 


96
GYTFTSYWLN
HCDR1;




Muc17.17,




Muc17.42,




Muc17.43,




Muc17.44,




Muc17.46,




Muc17.47


 


97
GYTFTSYWMN
HCDR1;




Muc17.4,




Muc17.5,




Muc17.6,




Muc17.15,




Muc17.16,




Muc17.18,




Muc17.19,




Muc17.20,




Muc17.32,




Muc17.33,




Muc17.34,




Muc17.35,




Muc17.36,




Muc17.37,




Muc17.38,




Muc17.39,




Muc17.40,




Muc17.41,




Muc17.45,




Muc17.48,




Muc17.49


 


98
GYTFTSYWMN
HCDR1;




Mcu17.31


 


99
MIHPSDSESRLNQKFKD
HCDR2;




Muc17.17.




Muc17.39,




Muc17.40,




Muc17.41,




Muc17.46,




Muc17.47


 


100
MIHPSDSETRLNQEFKD
HCDR2;




Muc17.5,




Muc17.20


 


101
MIHPSDSETRLNQKFKD
HCDR2;




Muc17.4,




Muc17.15,




Muc17.16,




Muc17.18,




Muc17.19,




Muc17.31,




Muc17.33,




Muc17.34,




Muc17.35,




Muc17.36,




Muc17.37,




Muc17.38,




Muc17.42,




Muc17.43,




Muc17.44,




Muc17.45,




Muc17.48,




Muc17.49


 


102
MIHPSDSETRLNQKFTD
HCDR2;




Muc17.6


 


103
QIYPGDGDINYNEKFRG
HCDR2;




Muc17.1,




Muc17.9,




Muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14,




Muc17.25,




Muc17.26,




Muc17.27,




Muc17.28,




Muc17.29,




Muc17.30,




Muc17.31


 


104
QIYPGDGDINYNGKFRG
HCDR2;




Muc17.3


 


105
QVYPGDDDINYNGKFRG
HCDR2;




Muc17.2


 


106
YISSGSSTIYYADTVKG
HCDR2;




Muc17.7,




Muc17.21,




Muc17.22,




Muc17.23,




Muc17.24


 


107
HGNYVMDY
HCDR2;




Muc17.8


 


108
HGNYVMDY
HCDR3;




Muc17.1,




Muc17.2,




Muc17.8,




Muc17.9,




Muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14,




Muc17.25,




Muc17.26,




Muc17.27,




Muc17.28,




Muc17.29,




Muc17.30,




Muc17.31


 


109
QGIITSVQEFAY
HCDR3;




Muc17.4,




Muc17.6,




Muc17.15,




Muc17.16,




Muc17.17,




Muc17.18,




Muc17.19,




Muc17.31,




Muc17.32,




Muc17.33,




Muc17.34,




Muc17.35,




Muc17.36,




Muc17.37,




Muc17.38,




Muc17.39,




Muc17.40,




Muc17.41,




Muc17.42,




Muc17.43,




Muc17.44,




Muc17.45,




Muc17.46,




Muc17.48,




Muc17.49


 


110
QGVITSVQEFAY
HCDR3;




Muc17.5,




Muc17.20


 


111
WGYYGSSYFAY
HCDR3;




Muc17.7,




Muc17.21,




Muc17.22,




Muc17.23,




Muc17.24


 


112
HGNYLMDY
HCDR3;




Muc17.3


 


113
SASSSLNYIY
LCDR1;




Muc17.6


 


114
SASSSVNYIF
LCDR1;




Muc17.18,




Muc17.19,




Muc17.41,




Muc17.44,




Muc17.46


 


115
SASSSVNYIY
LCDR1;




Muc17.4,




Muc17.5,




Muc17.15,




Muc17.16,




Muc17.17,




Muc17.31,




Muc17.32,




Muc17.33,




Muc17.34,




Muc17.35,




Muc17.36,




Muc17.37,




Muc17.38,




Muc17.39,




Muc17.40,




Muc17.41,




Muc17.43,




Muc17.45,




Muc17.47,




Muc17.48,




Muc17.49


 


116
SASSSVSYMF
LCDR1;




Muc17.1,




Muc17.2,




Muc17.8,




Muc17.9,




Muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14,




Muc17.25,




Muc17.26,




Muc17.27,




Muc17.28,




Muc17.29,




Muc17.30,




Muc17.31


 


117
SVSSNVDYVF
LCDR1;




Muc17.3


 


118
KASEDIYNRLA
LCDR1;




Muc17.7,




Muc17.21,




Muc17.22,




Muc17.23,




Muc17.24


 


119
RTSNLAS
LCDR2;




Muc17.1,




Muc17.2,




Muc17.4,




Muc17.5,




Muc17.6,




Muc17.8,




Muc17.9,




Muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14,




Muc17.15,




Muc17.16,




Muc17.17,




Muc17.18,




Muc17.19,




Muc17.20,




Muc17.25,




Muc17.26,




Muc17.27,




Muc17.28,




Muc17.29,




Muc17.30,




Muc17.31,




Muc17.32,




Muc17.33,




Muc17.34,




Muc17.36,




Muc17.37,




Muc17.38,




Muc17.39,




Muc17.40,




Muc17.41,




Muc17.42,




Muc17.43,




Muc17.44,




Muc17.45,




Muc17.46,




Muc17.48,




Muc17.49


 


121
GATNLET
LCDR2;




Muc17.7,




Muc17.21,




Muc17.22,




Muc17.23,




Muc17.24


 


122
QQFHDYPRT
LCDR3;




Muc17.1,




Muc17.8,




Muc17.9,




Muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14,




Muc17.25,




Muc17.26,




Muc17.27,




Muc17.28,




Muc17.29,




Muc17.30,




Muc17.31


 


123
QQFHSYPRT
LCDR3;




Muc17.2,




Muc17.3


 


124
QQFWRTPPT
LCDR3;




Muc17.7,




Muc17.21,




Muc17.22,




Muc17.23


 


125
QQYHSYPLT
LCDR3;




Muc17.4,




Muc17.5,




Muc17.6,




Muc17.15,




Muc17.16,




Muc17.17,




Muc17.18,




Muc17.19,




Muc17.20




Muc17.31,




Muc17.32,




Muc17.33,




Muc17.34,




Muc17.35,




Muc17.36,




Muc17.37,




Muc17.38,




Muc17.39,




Muc17.40,




Muc17.41,




Muc17.42,




Muc17.43,




Muc17.44,




Muc17.45,




Muc17.47,




Muc17.48,




Muc17.49


 


126
CQQFWRTPPT
LCDR3;




Muc17.24


 


127
EVQLVQSGAEVKKPGESLKISCKGSGYEFSSHWMNWVRQMPGK
VH;



GLEWMGQIYPGDGDINYNEKFRGQVTISADKSISTAYLQWSSLKA
Muc17.1,



SDTAMYYCARHGNYVMDYWGQGTLVTVSS
Muc17.8,




Muc17.9,




Muc17.10,




Muc17.11,




Muc17.12,




Muc17.13,




Muc17.14


 


128
QVQLVQSGAEVKKP+E60:J60GSSVKVSCKASGYTFTSYWLNWV
VH;



RQAPGQGLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYM
Muc17.17



ELSSLRSEDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


129
QVQLVQSGAEVKKPGASVKVSCKASGYEFSSHWMNWVRQAPGQ
VH;



GLEWMGQIYPGDGDINYNEKFRGRVTMTRDTSTSTVYMELSSLRS
Muc17.29,



EDTAVYYCARHGNYVMDYWGQGTLVTVSS
Muc17.30,




Muc17.31


 


130
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH



GLEWIGMIHPSDSETRLNQKFKDRVTLTVDKSSSTAYMELSSLRSE
Muc17.45



DTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


131
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQEFKDRVTMTRDTSTSTVYMELSSLR
Muc17.5,



SEDTAVYYCARQGVITSVQEFAYWGQGTLVTVSS
Muc17.20


 


132
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTLTRDKSISTAYMELSRLRS
Muc17.37



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


133
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH



GLEWMGMIHPSDSETRLNQKFKDRVTL TVDKSISTAYMELSRLRS
Muc17.35



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


134
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTLTVDTSISTAYMELSRLRS
Muc17.36



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


135
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTMTRDTSISTAYMELSRLRS
Muc17.23,



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS
Muc17.33,




Muc17.34


 


136
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTMTRDTSTSTVYMELSSLR
Muc17.31,



SEDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS
Muc17.48,




Muc17.49


 


137
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTMTVDKSISTAYMELSRLRS
Muc17.38



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


138
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
VH Muc17.6



GLEWMGMIHPSDSETRLNQKFTDRVTMTRDTSTSTVYMELSSLRS




EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS



 


139
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSDYWINWVRQAPGQ
VH Muc17.2



GLEWMGQVYPGDDDINYNGKFRGRVTITADKSTSTAYMELSSLRS




EDTAVYYCARHGNYVMDYWGQGTTVTVSS



 


140
QVQLVQSGAEVKKPGSSVKVSCKASGYEFSSHWMNWVRQAPGQ
Vh



GLEWMGQIYPGDGDINYNEKFRGRVTITADKSTSTA YMELSSLRS
Muc17.25,



EDTAVYYCARHGNYVMDYWGQGTTVTVSS
Muc17.26,




Muc17.27,




Muc17.28


 


141
QVQLVQSGAEVKKPGSSVKVSCKASGYIFSNHWMNWVRQAPGQ
VH;



GLEWMGQIYPGDGDINYNGKFRGRVTITADKSTSTAYMELSSLRS
Muc17.3



EDTAVYYCARHGNYLMDYWGQGTTVTVSS



 


142
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWLNWVRQAPGQ
VH;



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
Mucc. 17.46,



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS
Muc17.47


 


143
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWLNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
Muc17.42,



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS
Muc17.43,




Muc17.44


 


144
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
Muc17.39,



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS
Muc17.40,




Muc17.41


 


145
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
VH;



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
Muc17.4,



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSS
Muc17.15,




Muc17.16,




Muc17.18,




Muc17.19


 


146
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWVRQAPGK
VH;



GLEWVSYISSGSSTIYYADTVKGRFTISRDNAKNSLYLQMNSLRAE
Muc17.7,



DTAVYYCARWGYYGSSYFAYWGQGTLVTVSS
Muc17.21,




Muc17.22,




Muc17.23,




Muc17.24


 


147
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPKL
VL;



LIYGATNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQFW
Muc17.24



RTPPTFGGGTKVEIK



 


148
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPK
VL;



PLISGATNLETGVPSRFSGSGSGKDYTLTISSLQPEDIATYYCQQF
Muc17.21



WRTPPTFGGGTKVEIK



 


149
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPK
VL;



PLISGATNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQF
Muc17.7



WRTPPTFGGGTKVEIK



 


150
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPK
VL;



PLIYGATNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQF
Muc17.22



WRTPPTFGGGTKVEIK



 


151
EIVLTQSPATLSLSPGERATLSCSASSSVNYIFWYQQKPGQAPRLLI
VL;



YRTSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYHSY
Muc17.18



PLTFGGGTKVEIK



 


152
EIVLTQSPATLSLSPGERATLSCSASSSVNYIYWYQQKPGQAPRLLI
VL;



YRTSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYHSY
Muc17.16,



PLTFGGGTKVEIK
Muc17.17,




Muc17.34,




Muc17.39,




Muc17.42,




Muc17.49


 


153
EIVLTQSPATLSLSPGERATLSCSASSSVSYMFWYQQKPGQAPRL
VL;



LIYRTSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQFHD
Muc17.8,



YPRTFGGGTKVEIK
Muc17.27,




Muc17.31


 


154
EIVLTQSPATLSLSPGERATLSCSASSSVSYMFWYQQKPGQAPRL
VL;



LIYRTSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQFHS
Muc17.2



YPRTFGGGTKVEIK



 


155
EIVLTQSPATLSLSPGERATLSCSASSSVSYMFWYQQKPGQAPRP
VL;



WIYRTSNLASGIPPRFSGSGSGTDYTLTISSLEPEDFAVYYCQQFH
Muc17.28



DYPRTFGGGTKVEIK



 


156
EIVLTQSPATLSLSPGERATLSCSVSSNVDYVFWYQQKPGQAPRL
VL;



LIYRTSNLATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQFHS
Muc17.3



YPRTFGGGTKVEIK



 


157
EIVLTQSPATLSVSPGERATLSCSASSSVNYIYWYQQKPGQAPRP
VL;



WIYRTSNLASGIPARFSGSGSGTEYTLTISSLQSEDFAVYYCQQYH
Muc17.20



SYPLTFGGGTKVEIK



 


158
EIVMTQSPATLSVSPGERATLSCSASSSLNYIYWYQQKPGQAPRLL
VL;



IYRTSNLASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYHS
Muc17.6



YPLTFGGGTKVEIK



 


159
EIVMTQSPATLSVSPGERATLSCSASSSVNYIYWYQQKPGQAPRL
VL;



LIYRTSNLASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYHS
Muc17.5,



YPLTFGGGTKVEIK
Muc17.15,




Muc17.33,




Muc17.45,




Muc17.48


 


160
EIVMTQSPATLSVSPGERATLSCSASSSVSYMFWYQQKPGQAPRL
VL;



LIYRTSNLASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQFHD
Muc17.14,



YPRTFGGGTKVEIK
Muc17.26,




Muc17.30


 


161
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
VL;



YRTSNLASGVPPRFSGSGSGTEFTLTISSLQPEDFATYYCQQFHD
Muc17.13



YPRTFGGGTKVEIK



 


162
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
VL;



YRTSNLASGVPPRFSGSGSGTEYTLTISSLQPEDFATYYCQQFHD
Muc17.11



YPRTFGGGTKVEIKR



 


163
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
VL;



YRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQFHD
Muc17.9,



YPRTFGGGTKVEIK
Muc17.25,




Muc17.29


 


164
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
VL;



YRTSNLASGVPSRFSGSGSGTEYTLTISSLQPEDFATYYCQQFHD
Muc17.10



YPRTFGGGTKVEIK



 


165
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKPW
VL;



IYRTSNLASGVPPRFSGSGSGTEYTLTISSLQPEDFATYYCQQFHD
Muc17.12



YPRTFGGGTKVEIK



 


166
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKPW
VL;



IYRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQFHD
Muc17.1



YPRTFGGGTKVEIK



 


167
IQMTQSPSSLSASVGDRVTITCSASSSVNYIFWYQQKPGKAPKLLI
VL;



YRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYHS
Muc17.19,



YPLTFGGGTKVEIK
Muc17.41,




Muc17.44,




Muc17.46


 


168
IQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQQKPGKAPKLLI
VL;



YRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYHS
Muc17.4,



YPLTFGGGTKVEIK
Muc17.31,




Muc17.32,




Muc17.35,




Muc17.36,




Muc17.387,




Muc17.38,




Muc17.40,




Muc17.43,




Muc17.47


 


169
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPKL
VL;



LIYGATNLETGVPSRFSGSGSGKDYTLTISSLQPEDIATYYCQQFW
Muc17.23



RTPPTFGGGTKVEIK



 


170
GFTFSSFGMH
HCDR1;




CLDN182.3,




CLDN182.&,




CLDN182.12,




CLDN182.13


 


171
GYAFNNYWMN
HCDR1;




CLDN182.5


 


172
GYAFSSYWMN
HCDR1;




CLDN182.6,




CLDN182.9,




CLDN182.15,




CLDN182.16


 


173
GYTFTNFGIT
HCDR1;




CLDN182.4,




CLDN182.8,




CLDN182.10,




CLDN18.11


 


174
GYTFTNSGMN
HCDR1;




CLDN182.2,




CLDN182.14,




CLDN182.17


 


175
GYTFTNYGMN
HCDR1;




CLDN182.1


 


176
EIYPSSGNTFYNEKFKG
HCDR2;




CLDN182.4,




CLDN182.8,




CLDN182.10,




CLDN182.11


 


177
QISPGNGNSNFNGKFKG
HCDR2;




CLDN182.5


 


178
QIYPGNGNSNFNGKFKA
HCDR2;




CLDN182.6,




CLDN182.9,




CLDN182.15,




CLDN182.16


 


179
WINTNTGEPTFAEEFRG
HCDR2;




CLDN182.2,




CLDN182.14,




CLDN182.17


 


180
WINTNTGEPTYAEEFKG
HCDR2;




CLDN182.1


 


181
YISSGNSAIYYADTVNG
HCDR2;




CLDN182.3,




CLDN182.7,




CLDN182.12,




CLDN182.13


 


182
GGGPLRSRYFDY
HCDR3;




CLDN182.4,




CDLN182.8,




CLDN182.10,




CLDN182.11


 


183
GGRFGNAMDY
HCDR3;




CLDN182.6,




CLDN182.9,




CLDN182.15,




CLDN182.16


 


184
GGRYGNAMDY
HCDR3;




CLDN182.5


 


185
LRYGNSFDY
HCDR3;




CLDN182.3,




CLDN182.7,




CLDN182.12,




CLDN182.13


 


186
YFYGNSFVY
HCDR3;




CLDN182.1


 


187
YYYGNSFAY
HCDR3;




CLDN182.2,




CLDN182.14,




CLDN182.17


 


188
KSSQSLLNSGNQKAJYLT
LCDR1;




CLDN182.1,




CLDN182.2,




CLDN182.3,




CLDN182.6,




CLDN182.7,




CLDN182.9,




CLDN182.12,




CLDN182.13,




CLDN182.14,




CLDN182.15,




CLDN182.16,




CLDN182.17


 


189
KSSQSLLNSGNQRNYLT
LCDR1;




CLDN182.5


 


190
RSSQSLFSSGNQKNYLT
LCDR1;




CLDN182.4,




CLDN182.8,




CLDN182.10,




CLDN182.11


 


191
WVASTRES
LCDR2;




CLDN182.1,




CLDN182.2,




CLDN182.3,




CLDN182.4,




CLND182.5,




CLDN182.6,




CLDN182.7,




CLDN182.8,




CLDN182.9,




CLND182.10,




CLDN182.11,




CLDN182.12,




CLDN182.13,




CLDN182.14,




CLDN182.15,




CLDN182.16,




CLDN182.17


 


192
QNAYFYPYT
LCDR3;




CLDN182.5,




CLDN182.6,




CLND182.9,




CLDN182.15,




CLDN182.16


 


193
QNDYYYPLT
LCDR3;




CLDN182.4,




CLDN182.8,




CLND182.10,




CLDN182.11


 


194
QNNYFYPLT
LCDR3;




CLDN182.2,




CLND182.14,




CLND182.17


 


195
QNNYNFPLT
LCDR3;




CLDN182.1


 


196
QNNYYYPLT
LCDR3;




CLDN182.3,




CLND182.7,




CLND182.12,




CLND182.13


 


197
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHVWRQAPGK
VH;



GLEVWAYISSGNSAIYYADTVNGRFTISRDNPKNTLYLQMNSLRAE
CLDN182.12,



DTAVYYCARLRYGNSFDYWGQGTLVTVSS
CLND182.13


 


198
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHVWRQAPGK
VH;



GLEVWSYISSGNSAIYYADTVNGRFTISRDNAKNSLYLQMNSLRAE
CLDN182.3,



DTAVYYCARLRYGNSFDYWGQGTLVTVSS
CLDN182.7


 


199
QIQLVQSGAEVKKPGASVKVSCKASGYTFTNSGMNVWRQAPGQ
VH;



GLEWMGWINTNTGEPTFAEEFRGRVTFTLDTSASTAYMELSRLRS
CLDN182.14



DDTAVYYCARYYYGNSFAYWGQGTLVTVSS



 


200
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNFGITVWRQAPGQG
VH;



LEWIGEIYPSSGNTFYNEKFKGRVTLTADKSSSAAYMELRSLRSD
CLDN182.10,



DTAVYYCARGGGPLRSRYFDYWGQGTLVTVSS
CLND182.11


 


201
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNFGITVWRQAPGQG
VH;



LEWMGEIYPSSGNTFYNEKFKGRVTMTTDTSTSTAYMELRSLRSD
CLDN182.4,



DTAVYYCARGGGPLRSRYFDYWGQGTLVTVSS
CLND182.8


 


202
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNSGMNVWRQAPGQ
VH



GLEWMGWINTNTGEPTFAEEFRGRVTMTRDTSISTAYMELSRLRS
CLND182.2,



DDTAVYYCARYYYGNSFAYWGQGTLVTVSS
CLND182.17


 


203
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNVWRQAPGQ
VH;



GLEWMGWINTNTGEPTYAEEFKGRVTMTRDTSISTAYMELSRLRS
CLDN182.1



DDTAVYYCARYFYGNSFVYWGQGTLVTVSS



 


204
QVQLVQSGAEVKKPGSSVKVSCKASGYAFNNYWMNVWRQAPGQ
VH;



GLEWMGQISPGNGNSNFNGKFKGRVTITADKSTSTAYMELSSLRS
CLND182.5



EDTAVYYCARGGRYGNAMDYWGQGTTVTVSS



 


205
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSSYWMNVWRQAPGQ
VH;



GLEWIGQIYPGNGNSNFNGKFKARVTLTADKSSSTAYMELSSLRS
CLND182.15,



EDTAVYYCARGGRFGNAMDYWGQGTTVTVSS
CLND182.16


 


206
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSSYWMNVWRQAPGQ
VH;



GLEWMGQIYPGNGNSNFNGKFKARVTITADKSTSTAYMELSSLRS
CLDN182.6,



EDTAVYYCARGGRFGNAMDYWGQGTTVTVSS
CLDN182.9


 


207
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
VL;



PGQPPKLLIFWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND182.7,



YYCQNNYYYPLTFGGGTKVEIK
CLND182.13


 


206
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLDN182.6,



VYYCQNAYFYPYTFGGGTKVEIK
CLND182.15


 


207
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLND182.2,



VYYCQNNYFYPLTFGGGTKVEIK
CLND182.14




CLND182.17


 


208
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND182.1



YYCQNNYNFPLTFGGGTKVEIK



 


209
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLDN182.3,



VYYCQNNYYYPLTFGGGTKVEIK
CLND182.12


 


210
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQRNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLND182.5



VYYCQNAYFYPYTFGGGTKVEIK



 


211
DIVMTQSPDSLAVSLGERATINCRSSQSLFSSGNQKNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGADFTLTISSLQAEDVAV
CLDN182.8,



YYCQNDYYYPLTFGGGTKVEIK
CLND182.11


 


212
DIVMTQSPDSLAVSLGERATINCRSSQSLFSSGNQKNYLTWYQQK
VL;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLND812.4,



VYYCQNDYYYPLTFGGGTKVEIK
CLND182.10


 


213
DIVMTQSPDSLAVSLGERATMNCKSSQSLLNSGNQKNYLTWYQQ
VL;



KPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLND182.9,



VYYCQNAYFYPYTFGGGTKVEIK
CLND182.16


 


214
SGGGGS
1xG4S


 


215
GGGGSGGGGSGGGGS
3xG4S


 


216
GKPGSGKPGSGKPGSGKPGS
CL


 


217
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN




TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS




RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS




TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR




EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


218
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(N297A)



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS




RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAS




TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR




EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


219
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(L234F/L23



TKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMIS
5E/P331S)



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS




TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPR




EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


220
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(Y349C/K370E/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
K409D/K439E)



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS




TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR




EPQVCTLPPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALH




NHYTQESLSLSPGK



 


221
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(S354C/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
D356K/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
E357K/



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
D399K)



EPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


222
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(S354C/T366W)



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS




RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS




TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR




EPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


223
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(Y349C/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
T366S/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
L368A/



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
Y407V)



EPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


224
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(L234F/L23



TKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMIS
5E/P331S/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
Y349C/K37



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPR
0E/K409D/K



EPQVCTLPPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPEN
439E)



NYKTTPPVLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALH




NHYTQESLSLSPGK



 


225
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(L234F/L23



TKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMIS
5E/P331S/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
S354C/



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPR
D356K/



EPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN
E357K/



NYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH
0399K)



NHYTQKSLSLSPGK



 


226
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(L234F/L23



TKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMIS
5E/P331S/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
S354C/T36



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPR
6W)



EPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


227
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(L234F/L23



TKVDKRVEPKSCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMIS
5E/P331S/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS
Y349C/



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPR
T366S/



EPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPEN
L368A/



NYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALH
Y407V)



NHYTQKSLSLSPGK



 


228
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(N297A/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
Y349C/K37



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAS
0E/K409D/K



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
439E)



EPQVCTLPPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALH




NHYTQESLSLSPGK



 


229
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(N297A/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
S354C/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAS
D356K/



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
E357K/



EPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN
0399K)



NYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


230
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(N297A/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
S354C/T36



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAS
6W)



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR




EPQVYTLPPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


231
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
CH1-3 IgG1



LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
(N297A/



TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS
Y349C/



RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYAS
T366S/



TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR
L368A/



EPQVYTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPEN
Y407V)



NYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


232
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV




LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL




PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP




VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS




LSLSPGK



 


233
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSV
(N297A)



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL




PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP




VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS




LSLSPGK



 


234
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(L234F/L23



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTL
5E/P331S)



PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP




VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS




LSLSPGK



 


235
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(Y349C/K37



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTL
0E/K409D/K



PPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPP
439E)



VLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQES




LSLSPGK



 


236
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(S354C/



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
D356K/



PPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP
E357K/



VLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
D399K)



LSLSPGK



 


237
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(S354C/T36



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
6W



PPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPP




VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS




LSLSPGK



 


238
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(Y349C/



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
T366S/



PPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPP
L368A/



VLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
Y407V)



LSLSPGK



 


239
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(L234F/L23



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTL
5E/P331S/



PPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPP
Y349C/K37



VLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQES
0E/K409D/K



LSLSPGK
439E)


 


240
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(L234F/L23



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTL
5E/P331S/



PPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP
S354C/



VLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
D356K/



LSLSPGK
E357K/




D399K)


 


241
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(L234F/L23



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTL
5E/P331S/



PPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPP
S354C/T36



VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
6W)



LSLSPGK



 


242
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
(L234F/L23



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTL
5E/P331S/



PPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPP
Y349C/



VLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
T366S/



LSLSPGK
L368A/




Y407V)


 


243
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSV
(N297A/



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTL
Y349C/K37



PPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPP
0E/K409D/K



VLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQES
439E)



LSLSPGK



 


244
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSV
(N297A/



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
S354C/



PPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP
D356K/



VLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
E357K/



LSLSPGK
D399K)


 


245
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSV
(N297A/



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
S354C/T36



PPSRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPP
6W)



VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS




LSLSPGK



 


246
EPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
Fc IgG1



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSV
(N297A/



LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL
Y349C/



PPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPP
T366S/



VLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
L368A/



LSLSPGK
Y407V)


 


247
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN
CL1



ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT




HQGLSSPVTKSFNRGEC



 


248
EVQLVESGGGLVQPKGSLKLSCAASGFTFNTYAMNWVRQAPGKG
VH; CD3vA



LEWVARIRSKYNNYATYYADSVKDRFTISRDDSQSILYLQMNNLKT




EDTAMYYCVRHGNFGNSYVSWFAYWGQGTLVTVSS



 


249
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vB



GLEWVGRIRSKANNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


250
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vC



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


251
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vD



GLEWVGRIRSKINNYATYYADSVKGRFTISRDDSKNTLYLQMNSLR




AEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


252
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vE



GLEWVGRIRSKLNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


253
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vF



GLEWVGRIRSKVNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


254
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vG



GLEWVGRIRSKSNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


255
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMDWVRQAPGK
VH; CD3vH



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


256
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vI



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTVSS



 


257
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vJ



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFDYWGQGTLVTVSS



 


258
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vK



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSWFNYWGQGTLVTVSS



 


259
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
VH; CD3vL



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL




RAEDTAVYYCVRHGNFGDSYVSYFAYWGQGTLVTVSS



 


260
QLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLE
VH;



WVGRIRSKANNYATYYADSVKGRFTISRDDSKNTLYLQMNSLRAE
CD3vB2



DTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSS



 


261
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGK
VL; CD3



SPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYY




CALWYSNHWVFGGGTKLTVL



 


262
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vB



GLEWVGRIRSKANNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
scfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


263
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vC



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
scfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


264
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vD



GLEWVGRIRSKINNYATYYADSVKGRFTISRDDSKNTLYLQMNSLR
scfv-Fc



AEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSGK




PGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTT




SNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAAL




TISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHTC




PPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE




VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG




KEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQ




VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLY




SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


265
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vE



GLEWVGRIRSKLNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
scfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


266
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vF scfv-



GLEWVGRIRSKVNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
fc



RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


267
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vG



GLEWVGRIRSKSNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
scfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


268
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMDWVRQAPGK
CD3vH



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
scfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


269
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vI scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
Fc



RAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


270
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vJ scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
Fc



RAEDTAVYYCVRHGNFGDSYVSWFDYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


271
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vK scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
Fc



RAEDTAVYYCVRHGNFGDSYVSWFNYWGQGTLVTVSSGKPGSG




KPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVT




TSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAA




LTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTK




NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFF




LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


272
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3vI scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
Fc



RAEDTAVYYCVRHGNFGDSYVSYFAYWGQGTLVTVSSGKPGSGK




PGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTT




SNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAAL




TISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHTC




PPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE




VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG




KEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQ




VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLY




SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


273
QLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLE
CD3vB2



WVGRIRSKANNYATYYADSVKGRFTISRDDSKNTLYLQMNSLRAE




DTAVYYCVRHGNFGDSYVSWFAYWGQGTLVTVSSGKPGSGKPG




SGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSN




YANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTIS




GAQPEDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPP




CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK




FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE




YKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVS




LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKL




TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


274
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC; DLL3



GLEWVAVISGSGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA




EDTAVYYCARDWFFYLFDYWGQGTLVTVSSASTKGPSVFPLAPSS




KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


275
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC;



GLEWVAVISGSGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.11,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.15



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


276
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC:



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.18,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.22



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


277
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC; DLL3.5,



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.12,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.26



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


278
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC;



GLEWVAVISYDGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.19,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.23



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


279
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC;



GLEWVAVISYDGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.13,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.16,



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


280
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC;



GLEWVSVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.20,



EDTAVYYCAKDWFFYLFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.24



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


281
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
HC;



GLEWVSVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.14,



EDTAVYYCAKDWFYFIFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.17



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


282
EVQLQQSGPVLVKPGASVKMSCKASGFTFTDYYMNWVKQSHGK
HC;



SLEWIGVINPDNGITTYNQKFKGKATLTVDKSSSTAYMELNGLTSE
DLL3.21,



DSAVYYCARGVWNYERSFDYWGQGTTLTVSSASTKGPSVFPLAP
DLL3.25



SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ




SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC




DKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV




SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


283
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGK
HC; DLL3.9



GLEWVAVISHHGSSKYYADSVKGRYTISRDNSKNTLYLQMNSLRA




EDTAVYYCARDWFFYLFDYWGQGTLVTVSSASTKGPSVFPLAPSS




KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


284
QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGL
HC;



EWIGYVYYSGTTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTA
DLL3.27



VYYCASIAVTGFYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKST




SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY




SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT




CPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTK




NQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF




LYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


285
QVQLQQSGPVLVKPGASVKMSCKASGYSFTDYYVNWVKQSHGK
HC;



SLEWIGIISPNDGGTNYNQKFKGKATLTVDKSSSTAYMEVNSLTSE
DLL3.28



DSAVYYCARDDDLGWYFDVWGTGTTVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


286
QVQLVESGGGAVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
HC;



GLEWVAVISHHGSSKYYARSVKGRFTISRDNSKNTLYLEMNSLRA
DLL3.8



EDTAVYYCARDWWELVFDYWGQGTLVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


287
QVQLVESGGGAVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
HC;



GLEWVAVISHHGSSKYYARSVKGRFTISRDNSKNTLYLEMNSLRA
DLL3.29,



EDTAVYYCARDWWELVFDYWGQGTLVTVSSASTKGPSVFPLAPS
DLL3.30,



SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS
DLL3.36



SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


288
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
HC



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.10,



EDTAVYYCARDWFFYLFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.31



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


289
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGK
HC;



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
DLL3.32,



EDTAVYYCARDWFYFIFDYWGQGTLVTVSSASTKGPSVFPLAPSS
DLL3.33



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


290
QVQLVQSGAEVKKPGASVKVSCKASDYTFSNYYIEWVRQAPGQG
HC; DLL3.1



LEWMGEILPGNGNTVYNEKFKDRVTMTVDTSTSTAYMELRSLRSD




DTAVYYCARWGDYALFANWGQGTLVTVSSASTKGPSVFPLAPSS




KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELK




NQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF




LYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


291
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
HC; DLL3.3



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS




EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSASTKGPSVFP




LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA




VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK




SCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVV




DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV




LHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPP




SRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVL




DSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLS




LSPGK



 


292
QVQLVQSGAEVKKPGASVKVSCKATDYIFSNYYIEWVRQAPGQGL
HC; DLL3.2,



EWMGEILPGTGNTVYNEKFKDRVTMTVDTSTSTVYMELSSLRSED
DLL3.4



TAVYYCARWGDYALFANWGQGTLVTVSSASTKGPSVFPLAPSSK




STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG




LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH




TCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTK




NQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF




LYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


293
SDVQLQESGPGLVKPSQSLSLTCSVTGYSITSAYYWNWIRQFPGN
HC;



KLEWMGYISDVGHNYYNPSLKNRISITRDTSKNQFFLKLNSVTPED
DLL3.34



TATYYCARDQVFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSG




GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL




SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCP




PCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV




KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK




EYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQV




SLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS




DLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


294
SDVQLQESGPGLVKPSQSLSLTCSVTGYSITSAYYWNWIRQFPGN
HC;



KLEWMGYISDVGSNNYNPSLKNRISITRDTFKNQFFLKLNSVTTED
DLL3.35



TATYFCTRDQVFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSG




GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL




SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCP




PCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV




KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK




EYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTKNQV




SLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS




DLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


295
DIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQKPGKAPKL
LC; DLL3.3



LIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYS




TYPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF




YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


296
DIQMTQSPSFLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPK
LC; DLL3.4



PLIYSTSYRYSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQY




NNYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


297
DIQMTQSPSSLSASVGDRVTITCKSSQSLLHSDAKTFLYWYQQKP
LC;



GKAPKLLIYEVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATY
DLL3.26



YCLQGERLPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV




VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS




STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


298
DIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAPK
LC;



ALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQY
DLL3.27



NSYPFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


299
DIVMTQAAFSNPVTVGTSASISCRSSKSLLHSNGITYLYWYLQKPG
LC; DLL3.1



QSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVY




YCAQNLELPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV




VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS




STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


300
DIVMTQSPKFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQSP
LC; DLL3.2



KALIYSASYRYSGVPDRFTGSGSGTDFTLTFSSVQSEDLAEYFCQ




QYNNYPLTFGGGTKLEIKRRTVAAPSVFIFPPSDEQLKSGTASVVC




LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL




TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


301
DIVMTQSPLSLPVTPGEPASISCKSSQSLLHSDAKTFLYWYLQKPG
LC; DLL3.8



QSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY




CLQGRELPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV




CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS




TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


302
DIVMTQSPLSLPVTPGEPASISCKSSQSLLHSDGKTFLYWYLQKPG
LC; DLL3.9



QSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY




CLQGERLPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV




CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS




TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


303
DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSDAKTFLDWYLQKPG
LC; DLL3.22,



QSPQLLIYEVSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.23,



CLQGRELPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV
DLL3.24,



CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS
DLL3.25



TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


304
DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSDGKTFLDWYLQKPG
LC; DLL3.5,



QSPQLLIYEVSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.15,



CLQGERLPFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV
DLL3.16,



CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS
DLL3.17



TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


305
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDAKTFLYWYLQKPG
LC



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.18,



CLQGRELPFTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV
DLL3.19,



CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS
DLL3.20,



TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
DLL3.21


 


306
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDAKTFLYWYLQKPG
LC;



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.11,



CLQGRELPFTFGPGTKVEIKSTFGQGTKVEIKRTVAAPSVFIFPPSD
DLL3.12,



EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ
DLL3.13,



DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR
DLL3.14



GEC



 


307
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPG
LC



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.32,



CLQGERLPFTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV
DLL3.33



CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS




TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


308
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPG
LC;



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.10,



CLQGERLPFTFGPGTKVEIKSTFGQGTKVEIKRTVAAPSVFIFPPSD
DLL3.31



EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ




DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR




GEC



 


309
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPG
LC;



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.29,



CLQGIHLPFTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC
DLL3.30,



LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL
DLL3.36



TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


310
DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTFLYWYLQKPG
LC;



QPPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYY
DLL3.29.1



CLQGIHLPFTFGPGTKVEIKSTFGQGTKVEIKRTVAAPSVFIFPPSD




EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ




DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR




GEC



 


311
DTVLTQSPASLAVSLGQRATISCRASESVHSYGNSLIHWYQQKPG
LC;



QPPRLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYC
DLL3.34



QQTNEDPLTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVC




LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL




TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


312
EIVLTQSPGTLSLSPGERVTLSCRASQRVNNNYLAWYQQRPGQAP
LC;



RLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQ
DLL3.28



YDRSPLTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


313
QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKR
LC;



WIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQW
DLL3.35



SSNPLTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


341
QVQLVQSGAEVKKPGASVKVSCKASDYTFSNYYIEWVRQAPGQG
DLL3Scfv-



LEWMGEILPGNGNTVYNEKFKDRVTMTVDTSTSTAYMELRSLRSD
CD3Scfv-Fv



DTAVYYCARWGDYALFANWGQGTLVTVSSGGGGSGGGGSGGG




GSDIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGK




APKALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC




QQYNSYPFTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGSLRL




SCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYAD




SVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDSYV




SWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQE




PSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGG




TNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALWYSNH




WVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPK




DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE




EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKA




KGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESN




GQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVM




HEALHNHYTQKSLSLSPGK



 


342
QVQLVQSGAEVKKPGASVKVSCKATDYIFSNYYIEWVRQAPGQGL
DLL3Scfv-



EWMGEILPGTGNTVYNEKFKDRVTMTVDTSTSTVYMELSSLRSED
CD3Scfv-Fv



TAVYYCARWGDYALFANWGQGTLVTVSSGGGGSGGGGSGGGG




SDIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAP




KALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQ




YNSYPFTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGSLRLSCA




ASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSVK




GRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSW




FAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPS




LTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTN




KRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALWYSNHW




VFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKD




TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE




QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAK




GQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNG




QPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH




EALHNHYTQKSLSLSPGK



 


343
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3Scfv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
CD3Scfv-Fv



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG




SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNHYTQKSLSLSPGK



 


344
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3Scfv-



CLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
CD3Scfv-Fv



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG




SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGCGTKLEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNHYTQKSLSLSPGK



 


345
QVQLVQSGAEVKKPGASVKVSCKATDYIFSNYYIEWVRQAPGQGL
DLL3Scfv-



EWMGEILPGTGNTVYNEKFKDRVTMTVDTSTSTVYMELSSLRSED
CD3Scfv-Fv



TAVYYCARWGDYALFANWGQGTLVTVSSGGGGGGGGSGGGG




SDIQMTQSPSFLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAP




KPLIYSTSYRYSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQ




YNNYPLTFGGGTKVEIKSGGGSEVQLVESGGGLVQPGGSLRLSC




AASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSV




KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVS




WFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEP




SLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGT




NKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALWYSNH




WVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPK




DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE




EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKA




KGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESN




GQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVM




HEALHNHYTQKSLSLSPGK



 


346
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3Scfv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
CD3Scfv-Fv



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG




SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNHYTQKSLSLSPGK



 


347
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3Scfv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
CD3Scfv-Fv



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG




SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNHYTQKSLSLSPGK



 


348
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3Scfv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
CD3Scfv-Fv



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG




SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFNYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNHYTQKSLSLSPGK



 


349
EVQLVQSGAEVKKPGESLKISCKGSGYEFSSHWMNWVRQMPGK
Muc17scfv-



CLEWMGQIYPGDGDINYNEKFRGQVTISADKSISTAYLQWSSLKAS
CD3scfv



DTAMYYCARHGNYVMDYWGQGTLVTVSSGGGGSGGGGSGGGG




SGGGGSIQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPG




KAPKPWIYRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYY




CQQFHDYPRTFGCGTKVEIKSGGGSEVQLVESGGGLVQPGGSLR




LSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYA




DSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDSY




VSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQ




EPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG




GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALWYSN




HWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKP




KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR




EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISK




AKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWES




NGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSV




MHEALHNHYTQKSLSLSPGK



 


350
EVQLVQSGAEVKKPGESLKISCKGSGYEFSSHWMNWVRQMPGK
Muc17scfv-



GLEWMGQIYPGDGDINYNEKFRGQVTISADKSISTAYLQWSSLKA
CD3scfv



SDTAMYYCARHGNYVMDYWGQGTLVTVSSGGGGSGGGGSGGG




GSGGGGSIQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKP




GKAPKPWIYRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATY




YCQQFHDYPRTFGGGTKVEIKSGGGSEVQLVESGGGLVQPGGSL




RLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYY




ADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDS




YVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVT




QEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLI




GGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALWYS




NHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPK




PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP




REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTIS




KAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWE




SNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCS




VMHEALHNHYTQKSLSLSPGK



 


351
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKC
Muc17scfv-



LEWIGDIDASGSTKYNPSLKSRVTISLDTSKNQFSLKLNSVTAADTA
CD3scfv



VYFCARKKYSTVWSYFDNWGQGTLVTVSSGGGGSGGGGSGGG




GSSYELTQPSSVSVPPGQTASITCSGDKLGDKYASWYQQKPGQS




PVLVIYQDRKRPSGVPERFSGSNSGNTATLTISGTQAMDEADYYC




QAWGSSTAVFGCGTKLTVLSGGGGSEVQLVESGGGLVQPGGSL




KLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYY




ADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNS




YISYWAYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSL




TVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTK




FLAPGTPARFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRW




VFGGGTKLTVLGGGGDKTHTCPPCPAPELLGGPSVFLFPPKPKDT




LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPCEEQ




YGSTYRCVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG




QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ




PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE




ALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSGGGGSGGGGS




GGGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT




CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPCEEQYGSTYRCVS




VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT




LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP




PVLDSDGSFFLYSKLTVDKSRWQQGNVF




SCSVMHEALHNHYTQKSLSLSPGK



 


352
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNHGMHWVRQAPGK
Muc17scfv-



CLEWVAGIWSEGSNKYYADAVKGRFTISRDNSKNTLYLQMNSLRA
CD3scfv



EDTAVYYCARATYTTGWSYFDYWGQGTLVTVSSGGGGSGGGGS




GGGGSSYELTQPPSVSVSPGQTASITCSGDKLGDKYASWYQQKS




GQSPV




LVIYQDAKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQAF




HQSTWVFGCGTQLT




VLSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMN




WVRQAPGKGLEWVARIR




SKYNNYATYYADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYY




CVRHGNFGNSYISYW




AYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSPG




GTVTLTCGSSTGAVTSG




NYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGKAALTL




SGVQPEDEAEYYCVL




WYSNRWVFGGGTKLTVLGGGGDKTHTCPPCPAPELLGGPSVFLF




PPKPKDTLMISRTPEV




TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPCEEQYGSTYRCV




SVLTVLHQDWLNGKEY




KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL




TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL




TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSG




GGGSGGGGSGGGGSGGGGSGGGGSDKTHTCPPCPAPELLGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPCEEQYGSTYRCVSVLTVLHQDWLNGKEYKCKVSNKA




LPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


353
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWLNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIFWYQQ




KPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQPGG




SLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYAT




YYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG




DSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAV




VTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPR




GLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCAL




WYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFL




FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA




KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASI




EKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


354
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWLNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIFWYQQ




KPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQQYHSYPLTFGGGTKVEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNHYTQKSLSLSPGK



 


355
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWLNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQP




GGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNY




ATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGN




FGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQ




AVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKS




PRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYC




ALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPGK



 


356
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWbNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIfWYQQ




KPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQPGG




SLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYAT




YYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG




DSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAV




VTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPR




GLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCAL




WYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFL




FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA




KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASI




EKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


357
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWbNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQP




GGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNY




ATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGN




FGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQ




AVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKS




PRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYC




ALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPGK



 


358
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIfWYQQ




KPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQPGG




SLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYAT




YYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG




DSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAV




VTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPR




GLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCAL




WYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFL




FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA




KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASI




EKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPG



 


359
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSESRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQP




GGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNY




ATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGN




FGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQ




AVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKS




PRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYC




ALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPGK



 


360
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIfWYQQ




KPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQPGG




SLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYAT




YYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG




DSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAV




VTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPR




GLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCAL




WYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFL




FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA




KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASI




EKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


361
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfV-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGGSEVQLVESGGGLVQP




GGSLRLSCAASGFTFSTYAMNWVRQAFGKGLEWVGRIRSKYNNY




ATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGN




FGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQ




AVVTQERSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKS




PRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYC




ALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVQVSHEDPEVKFNWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPG



 


362
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTITISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGSEVQLVESGGGLVQPG




GSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWWGRIRSKINNYAT




YYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG




DSYVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAV




VTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPR




GLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCAL




WYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFL




FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA




KTKPREEQYNSTYRVVSVLIVLHQDWLNGKEYKCKVSNKALPASI




EKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


363
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGSEVQLVESGGGLVQPG




GSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYA




TYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNF




GDSYVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQA




VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSP




RGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA




LWYSNHWWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVF




LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN




AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAS




IEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


364
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGGGGGGGGSG




GGGSGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGSEVQLVESGGGLVQPG




GSLRLSCAASGFTFSTYAMNWVRQARGKGLEWYGRIRSKYNNYA




TYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNF




GDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQA




VVTQERSLTVSPGGTVTLTCGSSTGAVTTSNYANWWVQQKPGKSP




RGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA




LWYSNHWWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVF




LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVAN




AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAS




IEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALMNNYTQKSLSLSPGK



 


365
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
CD3scfv



EDTAVYYCARQGIITSVQEFNYWGQGTLVTVSSGGGGSGGGGSG




GGGGGGGGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQ




QKPGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQYHSYPLTFGGGTKVEIKSGGGSEVQLVESGGGLVQPG




GSLRISCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYA




TYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNF




GDSYVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQA




VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSP




RGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA




LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVF




LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN




AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAS




IEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


366
EVQLVQSGAEVKKPGESLKISCKGSGYEFSSHWMNWVRQMPGK
Muc17scfv-



GLEWMGQIYPGQGDINYNEKFRGQVTISADKSISTAYLQWSSLKA
Fc



SDTAMYYCARHGNYVMDYWGQGTLVTVSSGKPGSGKPGSGKPG




SGKPGSIQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPG




KAPKPWIYRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYY




CQQFHDYPRTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPGK



 


367
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
Muc17scfv-



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
Fc



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSGKPGSGKPGSGK




PGSGKPGSIQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQQK




PGKAPKLLIYRTSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATY




YCQQYHSYPLTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPS




VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV




HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL




PASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYP




SDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQ




GNVFSCSVMHEALHNHYTQKSLSLSPGK



 


368
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWWRQARGK
HC;



GLEWVSYISSGSSTIYYADTVKGRFTISRDNAKNSLYLQMNSLRAE
Muc17.7,



DTAVYYCARWGYYGSSYFAYWGQGTLVTVSSASTKGPSVFPLAP
Muc17.21,



SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ
Muc17.22,



SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSG
Muc17.23,



DKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV
Muc17.24



SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


369
EVQLVQSGAEVKKPGESLKISCKGSGYEFSSHWMNWVRQMPGK
HC;



GLEWMGQIYPGDGDINYNEKFRGQVTISADKSISTAYLQWSSLKA
Muc17.1,



SDTAMYYCARHGNYVMDYWGQGTLVTVSSASTKGPSVFPLAPSS
Muc17.8,



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS
Muc17.9.



GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT
Muc17.10,



HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE
Muc17.11,



DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW
Muc17.12,



LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT
Muc17.13,



KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLQSDGSF
Muc17.14



FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


370
QVQLVQSGAEVKKPGASVKVSCKASGYEFSSHWMNWWVRQAPGQ
HC;



GLEWMGQIYPGDGDINYNEKFRGRVTMTRDTSTSTVYMELSSLRS
Muc17.29,



EDTAVYYCARHGNYVMDYWGQGTLVTVSSASTKGPSVFPLAPSS
Muc17.30,



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS
Muc17.31



GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DREVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


371
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWIGMIHPSDSETRLNQKFKDRVILTVQKSSSTAYMELSSLRSE
Muc17.45



DTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVQKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


372
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNQEFKDRVTMTRDTSTSTVYMELSSLR
Muc17.5,



SEDTAVYYCARQGVITSVQEFAYWGQGTLVTVSSASTKGPSVFPL
Muc17.20



APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV




LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKPNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


373
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNQKFKDRVTLTRDKSISTAYMELSRLRS
Muc17.37



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLA




PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL




QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEREGGPSVFLAPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


374
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNQKFKDRVTLTVDKSISTAYMELSRLRS
Muc17.35



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLA




PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL




QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMMEALMNHYTQESLSL




SPGK



 


375
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNGKPKDRVTLTVDTSISTAYMELSRLRS
Muc17.36



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLA




PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL




QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


376
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNQKFKDRVTMTRDTSISTAYMELSRLRS
Muc17.232,



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLA
Muc17.33,



PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL
Muc17.34



QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


377
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC:



GLEWMGMIHPSDSETRLNQKFKDRVTMTRDTSTSTVYMELSSLR
Muc17.31,



SEDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLA
Muc17.48,



PSSKSTSGGTAALGCLVKDYFREPVTVSWNSGALTSGVHTFPAVL
Muc17.49



QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEPEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


378
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNQKFKDRVTMTVDKSISTAYMELSRLRS
Muc17.38



DDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLA




PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL




QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKFNWYVDGVEVNNAKTKPREEQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


379
QVQLVQSGAEVKKPGSSVKVSCKASGYEFSSHWMNWVRQAPGQ
HC



GLEWMGQIYPGDGDINYNEKFRGRVTITADKSTSTAYMELSSLRS
Muc17.25,



EDTAVYYCARHGNYVMDYWGQGTTVTVSSASTKGPSVFPLAPSS
Muc17.26,



KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS
Muc17.27,



GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT
Muc17,28



HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKENWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLNQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


380
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYWMNWVRQAPGQ
HC;



GLEWMGMIHPSDSETRLNQKFKDRVTITADKSTSTAYMELSSLRS
Muc17.4,



EDTAVYYCARQGIITSVQEFAYWGQGTLVTVSSASTKGPSVFPLAP
Muc17.15,



SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ
Muc17.16,



SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC
Muc17.18,



DKTHTCPPCPAPEFEGGPSVFLAPPKPKDTLMISRTPEVTCVVVDV
Muc17.19



SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


381
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPKL
LC;



LIYGATNLETGVPSRFSGSGSGKDYTLTISSLQPEDIATYYCQQFW
Muc17,23



RTPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN




FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK




ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


382
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPKL
LC;



LIYGATNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQFW
Muc17.24



RTPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN




FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK




ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


383
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPK
LC;



PLISGATNLETGVPSRFSGSGSGKDYTLTISSLQPEDIATYYCQQF
Muc17.21



WRTPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSENRGEC



 


384
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPK
LC;



PLISGATNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQF
Muc17.7



WRTPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSENRGEC



 


385
DIQMTQSPSSLSASVGDRVTITCKASEDIYNRLAWYQQKPGKAPK
LC;



PLIYGATNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQF
Muc17.22



WRTPPTFGGGTKVEIKRTVAARSVFIFRPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


386
EIVLTQSPATLSLSPGERATLSCSASSSVNYIYWYQQKPGQAPRLLI
LC;



YRTSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYHSY
Muc17.16,



PLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
Muc17.17,



REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD
Muc17.34,



YEKHKVYACEVTHQGLSSPVTKSFNRGEC
Muc17.39,




Muc17.42,




Muc17.49


 


387
EIVLTQSPATLSLSPGERATLSCSASSSVSYMFWYQQKPGQAPRL
LC;



LIYRTSNLASGIRARFSGSGSGTDFTLTISSLEPEDFAVYYCQQFHD
Muc17.8.



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF
Muc17.27,



YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
Muc17.31



DYEKHKVYACEVTHQGLSSPVTKSFNRGEG



 


388
EIVLTQSPATLSLSPGERATLSCSASSSVSYMFWYQQKPGQAPRP
LC;



WIYRTSNLASGIPPRFSGSGSGTDYTLTISSLEPEDFAVYYCQQFH
Muc17.28



DYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN




FYPREAKVQWKVQNALQSGNSQESVTEQDSKDSTYSLSSTLILSK




ADYEKHKVYACEVTHQGLSSPVIKSFNRGEC



 


389
EIVLTQSPATLSVSPGERATLSCSASSSVNYIYWYQQKPGQAPRP
LC;



WYRTSNLASGIPARFSGSGSGTEYTLTISSLQSEDFAVYYCQQYH
Muc17.20



SYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVGLLNNF




YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


390
EIVMTQSPATLSVSPGERATLSCSASSSVNYIYWYQQKPGQAPRL
LC;



LIYRTSNLASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYHS
Muc17.5,



YPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY
Muc17.15,



PREAKVQWKVQNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
Muc17.33,



DYEKHKVYACEVTHQGLSSVTKSFNRGEC
Muc17.45,




Muc17.48


 


391
EIVMTQSPATLSVSPGERATLSCSASSSVSYMFWYQQKPGQAPRL
LC;



LIYRTSNLASGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQFHD
Muc17.14,



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF
Muc17.26,



YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
Muc17.30



DYEKNKVYACEVTHQGLSSPVTKSENRGEC



 


392
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
LC;



YRTSNLASGVPPRFSGSGSGTEFTLTISSLQPEDFATYYCQQFHD
Muc17.13



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVQLLNNF




YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


393
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
LC;



YRTSNLASGVPPRFSGSGSGTEYTLTISSLQPEDFATYYCQQFHD
Muc17.11



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNA




YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


394
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
LC:



YRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQFHD
Muc17.9,



YPRTFGGGTKVEIKRTVAARSVFIFPPSDEQLKSGTASVVQLLNNF
Muc17.25.



YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
Muc17.29



DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


396
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKLLI
LC:



YRTSNLASGVPSRFSGSGSGTEYTLTISSLQPEDFATYYCQQFHD
Muc17.10



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVQLLNNF




YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


397
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKPW
LC;



IYRTSNLASGVPPRFSGSGSGTEYTLTISSLQPEDFATYYCQQPHD
Muc17.12



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF




YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


398
IQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGKAPKPWV
LC:



IYRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQFHD
Muc17.1



YPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVQLLNNF




YPREAKVQWKVQNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKÅ




DYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


399
IQMTQSPSSLSASVGDRVTITCSASSSVNYIYWYQQKPGKAPKLLI
LC;



YRTSNLASGVPSRFSGSGSGTDATLTISSLQPEDFATYYCQQYHS
Muc17.4,



YPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTÅSVVQLLNNFY
Muc17.31,



PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA
Muc17.32,



DYEKHKVYACEVTHQGLSSPVTKSFNRGEC
Muc17.35,




Muc17.36,




Muc17,387,




Muc17.38,




Muc17.40,




Muc17.43,




Muc17.47


 


400
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWWVQQKPGK
LC; CD3



SPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYY




CALWYSNHWVFGGGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASV




VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS




STLTLSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC



 


401
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWVRQAPGK
HC;



GLEWVAYISSGNSAIYYADTVNGRFTISRDNPKNTLYLQMNSLRAE
CLDN182.12



DTAVYYCARLRYGNSFDYWGQGTLVTVSSASTKGPSVFPLAPSSK
CLND182.13



STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG




LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH




TCPPCPAPEFEGGPSVELFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKPNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTK




NQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLQSDGSFF




LYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


402
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWRQAPGK
HC



GLEWVSYISSGNSAIYYADTVNGRFTISRDNAKNSLYLQMNSLRAE
CLDN182.3,



DTAVYYCARLRYGNSFDYWGQGTLVTVSSASTKGPSVFPLAPSSK
CLDN182.7



STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG




LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH




TCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED




PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL




NGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELTK




NQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF




LYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


403
QIQLVQSGAEVKKPGASVKVSCKASGYTFTNSGMNWVRQAPGQ
HC;



GLEWMGWINTNTGEPTFAEEFRGRVTFTLQTSASTAYMELSRLRS
CLDN182.14



DDTAVYYCARYYYGNSFAYWGQGTLVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYPPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYIGNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


404
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNFGITWVRQAPGQG
HC;



LEWIGETYPSSGNTFYNEKFKGRVTLTADKSSSAAYMELRSLRSCD
CLDN182.10,



TAVYYCARGGGPLRSRYFDYWGQGTLVTVSSASTKGRSVFPLAP
CLND182.11



SSKSTSGGTAALGCLVKDYPPEPVTVSWNSGALTSGVHTFPAVLQ




SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC




DKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV




SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


405
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNFGITWVRQAPGQG
HC;



LEWMGEIYPSSQNTFYNEKFKGRVTMTTDTSTSTAYMELRSLRSD
CLDN182.4,



DTAVYYCARGGGPLRSRYFDYWGQGTLVTVSSASTKGPSVEPLA
CLND182.8



PSSKSTSGGTAALGCLVKDYPPEPVTVSWNSGALTSGVHTFPAVL




QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS




CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVD




VSHEDPEVKPNWYVDGVEVHNAKTKPREFQYNSTYRVVSVLTVL




HQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPS




RDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLD




SDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSL




SPGK



 


406
QVQLVQSGAFVKKPGASVKVSCKASGYTFTNSGMNWVRQAPGQ
HC;



GLEWMGWINTNTGERTFAEEFRGRVTMTRDTSISTAYMELSRLRS
CLND182.2,



DDTAVYYCARYYYGNSFAYWGQGTLVTVSSASTKGPSVFPLAPS
CLND182.17



SKSTSGGTAALGCLVKDYFPERVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




CSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


407
QVQLVQSGAFVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQ
HC;



GLEWMGWINTNTGEPTYAEEFKGRVTMTRDTSISTAYMELSRLRS
CLDN182.1



DDTAVYYCARYFYGNSFVYWGQGTLVTVSSASTKGPSVFPLAPSS




KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS




GLYSLSSVVTVPSSSIGTQTYICNVNHKPSNTKVDKRVEPKSCDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRDELT




KNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF




FLYSDLIVQKSRWQQGNVFSCSVMHEALHNHYTQESLSLSPGK



 


408
QVQLVQSGAEVKKPGSSVKVSCKASGYAFNNYWMNWVRQAPGQ
HC;



GLEWMGQISPGNGNSNFNGKFKGRVTITADKSTSTAYMELSSLRS
CLND182.5



EDTAVYYCARGGRYGNAMDYWGQGTTVTVSSASTKGPSVFPLAP




SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ




SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC




DKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV




SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


409
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSSYWMNWVRQAPGQ
HC;



GLEWIGQIYPGNGNSNFNGKFKARVTLTADKSSSTAYMELSSLRS
CLND182.15,



EDTAVYYCARGGRFGNAMDYWGQGTTVTVSSASTKGPSVFPLAP
CLND182.16



SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ




SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVQKRVEPKSG




DKTHTCPPCPAPEREGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV




SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


410
QVQLVQSGAEVKKPGSSVKVSCKASGYAFSSYWMNWWRQAPGQ
HC;



GLEWMGQIYPGNGNSNFNGKFKARVTITADKSTSTAYMELSSLRS
CLDN182.6,



EDTAVYYCARGGRFGNAMDYWGQGTTVTVSSASTKGPSVFPLAP
CLDN182.9



SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ




SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC




DKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV




SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH




QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSR




DELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDS




DGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLS




PGK



 


411
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
LC;



PGQPPKLLIFWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND182.7,



YYCQNNYYYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS
CLND182.13



VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


412
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLDN182.6,



YYCQNAYFYPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS
CLND182.15



VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


413
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND182.2,



YYCQNNYFYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS
CLND182.14,



VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL
GUND182.17



SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


414
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND182.1



YYCQNNYNFPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS




VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYAGEVTHQGLSSPVTKSFNRGEC



 


415
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLDN182.3,



YYCQNNYYYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS
CLND182.12



VVCLLNNFYPREAKVQWKVQNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


416
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQRNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND182.5



YYCQNAYFYPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS




VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


417
DIVMTQSPDSLAVSLGERATINGRSSQSLFSSGNQKNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGADFTLTISSLQAEDVAV
CLDN182.8,



YYCQNDYYYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS
CLND182.11



VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSENRGEC



 


418
DIVMTQSPDSLAVSLGERATINGRSSQSLFSSGNQKNYLTWYQQK
LC;



PGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV
CLND812.4,



YYCQNDYYYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTAS
CLND182.10



VVCLLNNFYPREAKVQWKVQNALQSGNSQESVTEQDSKDSTYSL




SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


419
DIVMTQSPDSLAVSLGERATMNCKSSQSLLNSGNQKNYLTWYQQ
LC;



KPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVA
CLND182.9,



VYYCQNAYFYPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTA
CLND182.16



SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYS




LSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


420
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
VH; CD28



LEWIGCIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD




TAVYFCTRSHYGLQWNFDVWGQGTTVTVSS



 


421
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
VH; CD28



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50S)



TAVYFCTRSHYGLDWNFDVWGQGTTVTVSS



 


422
QVQLVQSGAEVKKRGASVKVSCKASGYTFTSYYIHWVRQAPGQG
VH; CD28



LEWIGAIYPGNVNTNYNEKFKDRATLTVQTSISTAYMELSRLRSDD
(C50A)



TAVYFCTRSHYGLDWNFDVWGQGTTVTVSS



 


423
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
VH; CD28



LEWIGGIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50G)



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSS



 


424
DIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQQKPGKAPK
VL CD28



LLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQG




QTYPYTFGGGTKVEIK



 


425
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
HC; CD28



LEWIGCIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD




TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSASTKGPSVFPLARS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVERKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREFQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKITPPVLQSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


426
DIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQQKRGKAPK
LC; CD28



LUYKASNIHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQG




QTYPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN




NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS




KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


427
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
CD28 Scfv-



LEWIGCIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
Fc



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSGKPGSGKPGSGKP




GSGKPGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQ




QKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQGQTYPYTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


428
QVQLVQSGAFVKKPGASVKVSCKASGYTFTSYYIHWRQAPGQG
CD28



LEWIGGIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50G)



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSGKPGSGKPGSGKP
Scfv-Fc



GSGKPCSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQ




QKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQGQTYPYTFGGGTKVEIKERKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


429
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
CD28



LEWIGAIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50A) scfv-



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSGKPGSGKPGSGKP
Fc



GSGKPGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQ




QKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDA




ATYYCQQGQTYPYTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVWVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


430
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
CD28



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50S)



TAVYFCTRSHYGLDWNFDVWGQGTTVTVSSGKPGSGKPGSGKP
scfv-Fc



GSGKPGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQ




QKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTITISSLQPEDF




ATYYCQQGQTYPYTFGGGTKVEIKERKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDG




VEVHNAKTKPREEQYNSTYRVVSVILTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


431
QVQLVQSGAEVKKPGASVKVSCKASDYTFSNYYIEWVRQAPGQG
DLL3scfv-



LEWMGEILPGNGNTVYNEKFKDRVTMTVDTSTSTAYMELRSLRSD
Fc



DTAVYYCARWGDYALFANWGQGTLVTVSSGGGGGGGGSGGG




GSDIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGK




APKALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC




QQYNSYPFTFGQGTKLEIKERKSSDKTHTCPPCPAPEFEGGPSVF




LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN




AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAS




IEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALNNHYTQKSLSLSPGK



 


432
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3scfv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVQRSTSTVYMELSSLRS
Fc



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG




SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKERKSSDKTHTCPPCPAPEFEGGP




SVFLAPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVQKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


433
QVQLVQSGAEVKKPGASVKVSCKATDYIFSNYYIEWVRQAPGQGL
DLL3scfv-



EWMGEILPGTGNTVYNEKFKDRVTMTVDTSTSTVYMELSSLRSED
Fc



TAVYYCARWGDYALFANWGQGTLVTVSSGGGGGGGGSGGGG




SDIQMTQSPSFLSASVGDRVTITCKASQNVGTNVAWYQQKGKAP




KPLYSTSYRYSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQ




YNNYPLTFGGGTKVEIKERKSSDKTHTCPPCPAPEFEGGPSVFLFR




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMMEALMNMYTQKSLSLSPGK



 


434
QVQLVQSGAEVKKPGASVKVSCKATDYIFSNYYIEWWVRQAPGQGL
DLL3scfV-



EWMGEILPGTGNTVYNEKFKDRVTMTVDTSTSTVYMELSSLRSED
Fc



TAVYYCARWGDYALFANWGQGTLVTVSSGGGGGGGGGSGGGG




SDIQMTQSPSTLSASVGDRVTITCKASQNVGTNVAWYQQKPGKAP




KALIYSASYRYSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQ




YNSYPFTFGQGTKLEIKEPKSSDKTHTCFPCPAPEFEGGPSVFLFP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMHEALHNMYTQKSLSLSPGK



 


435
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQC
CD28



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50S)



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSGKPGSGKPGSGKP
Scfv(CC)-Fc



GSGKPGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQ




QKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQGQTYPYTFGCGTKVEIKERKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


436
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
CD28



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
(C50S)



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSGGGGGGGGSGG
Scfv(G4S)-



GGSGGGGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWY
Fc



QQKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPED




FATYYCQQGQTYPYTFGGGTKVEIKERKSSDKTHTCPPCPAREFE




GGPSVFLPPPKPKDTLMISRTPEVTGVVVDVSHEDPEVKPNWYVD




GVEVHNAKTKPREEQYNSTYRVVSVLTVLNQDWINGKEYKCKVS




NKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVK




GFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKS




RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


437
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
HC;



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
CD137.U



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSIGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


438
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LC;



LUIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRS
CD137.U



NWPPALTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL




NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT




LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


439
QVQLVQSGAEVKKPGASVKVSCKASGYTESSYWMHWVRQAPGQ
HC;



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
CD137.B



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSIGTQTYICNVNHKPSNTKVQKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTGVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTGLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP



 


440
DIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQKPGQSPR
LC;



LUKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQDGH
CD137.B



SFPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVQLLNN




FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK




ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


441
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
CD137.U



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
scfv(CL)-Fc



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGKPGSGKPGSGKP




SGKPGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQ




KPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQQRSNWPPALTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


442
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
CD137.U



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
scfv(G4S)-



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGGGGGGGGGGGG
Fc



GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ




APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ




QRSNWPPALTFGGGTKVEIKREPKSSDKTHTCPPCPAPEFEGGPS




VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV




HNAKTKPREEQYNSTYRVVSVLTVLHQDWINGKEYKCKVSNKAL




PASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYP




SDIAVEWESNGQPENNYKITPPVLKSDGSFFLYSKLTVQK




GNVFSCSVMHEALHNHYTQKSLSLSPGK



 


443
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
CD137.U



LEWIGEINHGGYVTYNPSLESRVTISVQTSKNQFSLKLSSVTAADTA
scfv (6x)-Fc



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGGGGGGGGGGGG




GGSGGGGGGGGSGGGGSEIVLTQSPATLSLSFGERATLSCRAS




QSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDF




TLTISSLEPEDFAVYYCQQRSNWPPALTFGGGTKVEIKEPKSSDKT




HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE




DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW




LNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLT




KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSF




FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


444
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
CD137.U



LUIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRS
scfv(VLVH)



NWPPALTFGGGTKVEIKGKPGSGKPGSGKPGSGKPGSQVQLQQ
Fc



WGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKGLEWIGEI




NHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTAVYYCAR




DYGPGNYDWYFDLWGRGTLVTVSSEPKSSDKTHTCPPCPAPEFE




GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD




GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS




NKALPASIEKTISKAKGQPREPQVYTLPPCRKKLIKNQVSLTCLVK




GFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKS




RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


445
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
CD137.U(G



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
99S) scfv-



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGGGGSGGGGSGG
Fc



GGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ




APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ




QRSNWPPALTFSGGTKVEIKREPKSSDKTHTCPFCPAPEFEGGPS




VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV




HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL




PASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYP




SDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQ




GNVFSCSVMHEALHNHYTQKSLSLSPGK



 


446
QVQLQESGPGLVKPSETLSLTCTVSGGSFSGYYWSWIRQPPGKG
CD137.Uv1



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
scfv-Fc



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGKPGSGKPGSGKP




GSGKPGSEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQ




QKPGQAPRLLIYDASNRATGIFDRFSGSGSGTDFTLTISRLEPEDF




AVYYCQQRSNWPPALTFGGGTKVEIKEPKSSDKTHTCPPCPAPEF




EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV




DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV




SNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLV




KGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDK




SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


447
EVQLLESGGGLVQPGGSLRLSCAASGGSFSGYYWSWVRQAPGK
CD137.Uv2



GLEWWVSEINHGGYVTYNPSLESRFTISRDNSKNTLYLQMNSLRAE
scfv-Fc



DTAVYYCAKDYGPGNYDWYFDLWGRGTLVTVSSGKPGSGKPGS




GKPGSGKPGSEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLA




WYQQKPGQAPRLLIYDASNRATGIPDRESGSGSGTDFTLTISRLER




EDFAVYYCQQRSNWPPALTFGGGTKVEIKEPKSSDKTHTCPPCPA




PEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKEN




WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK




CKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLT




CLVKGFYPSDIAVEWESNGQPENNYKTTPRVLKSDGSFFLYSKLT




VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


448
QVQLVQSGAEVKKPGSSVKVSCKASGGSFSGYYWSWVRQAPGQ
CD137.Uv3



GLEWMGEINHGGYVTYNPSLESRVTITADESTSTAYMELSSLRSE
scfv-Fc



DTAVYYCARDYGPGNYDWYFDLWGRGTLVTVSSGKPGSGKPGS




GKPGSGKPGSEIVLTQSPGTLSLSPGERATLSGRASQSVSSSYLA




WYQQKPGQAPRLLIYDASNRATGIPDRFSGSGSGTDFTLTISRLEP




EDFAVYYCQQRSNWPPALTFGGGTKVEIKEPKSSDKTHTCPPCPÅ




PEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKEN




WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK




CKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLT




CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLT




VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


449
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKC
CD137



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA




VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGKPGSGKPGSGKP




GSGKPGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQ




KPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQQRSNWPPALTFGCGTKVEIKEPKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


450
QVQLGQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKC
CD137.U



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
scfv(CC)-Fc



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSGKPGSGKPGSGKP




GSGKPGSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQ




KPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQQRSNWPPALTFGCGTKVEIKEPKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


451
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWRQAPGQ
CD137.B



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
scfv-Fc



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQ




KPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALMNHYTQKSLSLSPGK



 


452
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWRQAPGQ
CD137.b(S60A)



RLEWMGEINPGNGHTNYAQKFQGRVTITVDKSASTAYMELSSLRS
scfv-fc



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQ




KPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPÅPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALMNHYTQKSLSLSPGK



 


453
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQ
CD137.B



RLEWMGEINPGNGHTNYNQKFQGRVTITVDKSASTAYMELSSLRS
(S50N)



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKP
scfv-fc



GSGKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQ




KPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKPNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


454
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQAPGK
CD137.39



GLEWVADIKNDGSYTNYAPSLTNRFTISRDNAKNSLYLQMNSLRA
scfv-Fc



EDTAVYYCARELTGTWGQGTMVTVSSGKPGSGKPGSGKPGSGK




PGSDIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAWY




QQKPGQPPKLLIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAED




VAVYYCLQYDRYPFTFGQGTKLEIKEPKSSDKTHTCPPCPAPEFE




GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD




GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS




NKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVK




GFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKS




RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


455
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQAPGK
CD137.39v



GLEWVADIKNDGSYTNYVDSVKGRFTISRDNAKNSLYLQMNSLRA
1 scfv-Fc



EDTAVYYCARELTGTWGQGTLVTVSSGKPGSGKPGSGKPGSGKP




GSDIIMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAWYQ




QKPGQPPELLIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAEDV




AVYYCLQYDRYPFTEGQGTKLEIKERKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


456
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWWVRQAPGK
CD137.39v



GLEWVADIKNDGSYTNYVDSVKGRFTISRDNAKNSLYLQMNSLRA
2 scfv-Fc



EDTAVYYCARELTGTWGQGTLVTVSSGKPGSGKPGSGKPGSGKP




GSDIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAWYQ




QKPGQPPKLUIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAEDV




AVYYCLQYDRYPFTEGQGTKLEIKERKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


457
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQAPGK
CD137.39v



GMEWVGDIKNDGSYTNYAPSLTNRFTISRDNARNSLYLQMNSLRA
3 scfv-Fc



EDTAVYYCTRELTGTWGQGTLVTVSSGKPGSGKPGSGKPGSGKP




GSDIIMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAWYQ




QKPGQPPELLIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAEDV




AVYYCLQYDRYPFTFGQGTKLEIKERKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVQG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


458
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQAPGK
CD137.39V



GMEWVGDIKNDGSYTNYAPSLTNRFTISRDNARNSLYLQMNSLRA
4 scfv-Fc



EDTAVYYCTRELTGTWGQGTLVTVSSGKPGSGKPGSGKPGSGKP




GSDIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAWYQ




QKPGQPPKLUYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAEDV




AVYYCLQYDRYPFTFGQGTKLEIKEPKSSDKTHTCPPCPAPEFEG




GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG




VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN




KALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKG




FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR




WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


459
QVKLVESGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.Bv1



VLEWMGEINPGNGHTSYAQKFQGRVTLTVDKSTSTAYMELSSLRS
scfv-Fc



EDTAVYYCARSFTTARAFAYWGQGTTVTVSSGKPGSGKPGSGKP




GSCKPGSDIQMTQSPSSLSASVGDRVTITCRASQSISDYLHWYQQ




KPGKAPKLLIKYASQSISGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQDSHSFPPTFGGGTKVEIKERKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


460
QVKLVESGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.Bv2



VLEWMGEINPGNGHTSYAQKFQGRVTLTVDKSTSTAYMELSSLRS
scfv-fc



EDTAVYYCARSFTTARAFAYWGQGTTVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQ




KPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


461
QVQLVQSGAEVKKPGASVKLSCKASGYTFSSYWMEIWVRQAPGQ
CD137.Bv3



GLEMGEINPGNGHTNYNEKFKSRVTMTRDTSTSTAYMELSSLRS
scfv-fc



EDTAVYYCARSFKTARAFAYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPAFLSVTRGEKVTITCRASQTISDYLHWYQQ




KPDQAPKLUKYASQSISGIPSRFSGSGSGTDFTFTISSLEAEDAAT




YYCQDGHSWPPTFGQGTKLEIKEPKSSDKTHTCPPCPAPEFEGG




PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV




EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK




ALPASIEKTISKAKGGPREPQVYTLPPCRKKLTKNQVSLTCLVKGF




YPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRW




QQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


462
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.Bv4



GLEWMGIINPGNGHTSYAQKFQGRVTMTRDTSTSTVYMELSSLRS
scfv-fc



EDTAVYYCARSFTTARAFAYWGQGTTVTVSSGKPGSGKPGSGKP




GSGKPGSDIQMTQSPSSLSASVGDRVTITGRASQSISDYLHWYQQ




KPGKAPKLLIKYASQSISGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQDSHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


463
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.Bv5



GLEWMGIINPGNGHTSYAQKFQGRVTMTRDTSTSTVYMELSSLRS
scfv-fc



EDTAVYYCARSFTTARAFAYWGQGTTVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQ




KPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPÅPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


464
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.Bv6



RLEWMGEINPSNGHTKYSQKFQGRVTITVDKSASTAYMELSSLRS




EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIQMTQSPSSLSASVGDRVTITCRASQSISDYLHWYQQ




KPGKAPKLLIKYASQSISGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQDSHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


465
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.Bv7



RLEWMGEINPSNGHTKYSQKFQGRVTITVDKSASTAYMELSSLRS
scfv-fc



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQ




KPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAV




YYCQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


466
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3 scfv-



CLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
Fc



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGKPGSGKPG




SGKPGSGKPGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVA




WYQQKPGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQP




EDFATYYCQQYSTYPYTFGCGTKLEIKEPKSSGKTHTCPPCPAPER




EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYV




DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV




SNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLV




KGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDK




SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


467
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGK
DLL3 scfv-



GLEWVAVISHHGSSKYYADSVKGRFTISRDNSKNTLYLQMNSLRA
Fc



EDTAVYYCARDWFFYLFDYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPLSLPVTPGEPASISCKSSQSLLHSDGKTFLY




WYLQKPGQSPQLLIYEVSNRFSGVPDRFSGSGSGTDFTLKISRVE




AEDVGVYYCLQGERLPFTFGQGTKVEIKEPKSSDKTHTCPPCPAP




EFEGGRSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW




YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC




KVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTC




LVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTV




DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


468
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3scFv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
41BBscFv-fc x



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGSGGGG
41BBFab-fc



SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK




PCKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKSGGGGSQVQLVQSGAEVKKPGA




SVKVSCKASGYTFSSYWMHWRQAPGQRLEWMGEINPGNGHTN




YSQKFQGRVTITVDKSASTAYMELSSLRSEDTAVYYCARSFTTARA




FAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSDIVMTQSPPT




LSLSPGERVTLSCRASQSISDYLHWYQQKPGQSPRLLIKYASQSIS




GIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQDGHSFPPTFGGGT




KVEIKEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRT




PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY




RVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREP




QVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY




KTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH




YTQKSLSLSPGK



 


469
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQARGQ
4-1BBscFv-



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
41BBscFv-Fc X



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGGGGGGGGGGGG
DLL3Fab-Fc



GGSGGGGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQ




QKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFA




VYYCQDGHSFPPTFGGGTKVEIKSGGGGSQVQLVQSGAEVKKPG




ASVKVSCKASGYTFSSYWMHWVRQAPGQRLFWMGFINPGNGHT




NYSQKFQGRVTITVDKSASTAYMELSSLRSEDTAVYYCARSFTTA




RAFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSDIVMTQSP




PTLSLSPGERVTLSCRASQSISDYLHWYQQKPGQSPRLLIKYASQS




ISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQDGHSFPPTFGG




GTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMIS




RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS




TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPR




EPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN




NYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH




NHYTQKSLSLSPGK



 


470
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQ
41BBFab-



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
Fc-



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSASTKGPSVFPLAPS
DLL3scFv



SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GKSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMN




WVRQAPGQGLEWMGVINPYNDITIYNQKFQGRVTMTVQRSTSTV




YMELSSLRSEDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSG




GGGSGGGGSGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGI




AVAWYQQKPGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISS




LQPEDFATYYCQQYSTYPYTFGQGTKLEIK



 


471
QVQLQQWGAGLLKPSETLSLTCAVYGGSESGYYWSWIRQSPEKG
41BBFab-



LEWIGEINHGGYVTYNPSLESRVTISVQTSKNQFSLKLSSVTAADTA
Fc-



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSASTKGPSVFPLAPS
DLL3scFv



SKSTSGGTAALGCLVKDYFREPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRK




KLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSD




GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP




GKSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMN




WRQAPGQGLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTV




YMELSSLRSEDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSG




GGGSGGGGSGGGGSDIQITQSPSFLSASVGDRVTITCKASQNVGI




AVAWYQQKPGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISS




LQPEDFATYYCQQYSTYPYTFGQGTKLEIK



 


472
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQ
41BBFab-



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
DLL3scFv



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVERKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKPNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRK




KLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSD




GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP




GKSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMN




WVRQAPGQGLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTV




YMELSSLRSEDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSG




GGGGGGGGGGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGI




AVAWYQQKPCKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISS




LQPEDFATYYCQQYSTYPYTFGQGTKLEIK



 


473
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
41BBFab-



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
Fc-



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSASTKGPSVFPLAPS
DLL3scFv



SKSTSGGTAALGCLVKDYFPEFVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GKSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMN




WVRQAPGQGLEWMGVINPYNDITTYNQKFQGRVTMTVQRSTSTV




YMELSSLRSEDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSG




GGGGGGGSGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGI




AVAWYQQKPGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISS




LQPEDFATYYCQQYSTYPYTFGQGTKLEIK



 


474
EVQLVQSGAEVKKPGESLKISCKGSGYEFSSHWMNWVRQMPGK
Muc17scfv-



CLEWMGQIYPGDGDINYNEKFRGQVTISADKSISTAYLQWSSLKAS
fc



DTAMYYCARHGNYVMDYWGQGTLVTVSSGKPGSGKPGSGKPGS




GKPGSIQLTQSPSFLSASVGDRVTITCSASSSVSYMFWYQQKPGK




APKPWYRTSNLASGVPSRFSGSGSGTEFTLTISSLQPEDFATYYC




QQFHDYPRTFGCGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSVF




LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN




AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAS




IEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


475
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNFGITWVRQAPGQG
CLDN182.



LEWMGETYPSSQNTFYNEKFKGRVTMTTDTSTSTAYMELRSLRSD
scfv-Fc



DTAVYYCARGGGPLRSRYFDYWGQGTLVTVSSGKPGSGKPGSG




KPGSGKPGSDIVMTQSPDSLAVSLGERATINGRSSQSLFSSGNQK




NYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTI




SSLQAEDVAVYYCQNDYYYPLTFGGGTKVEIKEPKSSDKTHTCFP




CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK




FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE




YKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVS




LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKL




TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


476
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGMHWWRQAPGK
CLDN182.



GLEWWSYISSGNSAIYYADTVNGRFTISRDNAKNSLYLQMNSLRAE
scfv-Fc



DTAVYYCARLRYGNSFDYWGQGTLVTVSSGKPGSGKPGSGKPG




SGKPGSDIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYL




TWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSL




QAEDVAVYYCQNNYYYPLTFGGGTKVEIKEPKSSDKTHTCPPGRA




PEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKEN




WYVDGVEVHNAKTKPREFQYNSTYRVVSVLTVLHQDWLNGKEYK




CKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLT




CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLT




VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


477
QVQLVQSGAEVKKPGSSVKVSCKASGYAFNNYWMNWVRQAPGQ
CLDN182.



GLEWMGQISPGNGNSNFNGKFKGRVTITADKSTSTAYMELSSLRS
scfv-Fc



EDTAVYYCARGGRYGNAMDYWGQGTTVTVSSGKPGSGKPGSGK




PGSGKPGSDIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQRN




YLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTIS




SLQAEDVAVYYCQNAYFYPYTFGGGTKVEIKEPKSSDKTHTCPPC




PAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF




NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY




KCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSL




TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLT




VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


478
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNSGMNWVRQAPGQ
CLDN182.



GLEWMGWINTNTGEPTFAEEFRGRVTMTRDTSISTAYMELSRLRS
scfv-Fc



DDTAVYYCARYYYGNSFAYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSDIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNY




LTWYGQKPGQPPKLUYWASTRESGVPDRFSGSGSGTDFTLTISS




LQAEDVAVYYCQNNYFYPLTFGGGTKVEIKEPKSSDKTHTCPPCP




APEFEGGPSVFLPPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN




WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK




CKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLT




CLVKGFYPSDIAVEWESNGQPENNYKITPPVLKSDGSFFLYSKLT




VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


479
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMHWVRQAPGQ
CD137.bV8



GLEWMGIINPGNGHTSYAQKFQGRVTMTRDTSTSTVYMELSSLRS
scfv-Fc



EDTAVYYCARSFTTARAFAYWGQGTTVTVSSGKPGSGKPGSGKP




GSGKPGSDIQMTQSPSSLSASVGDRVTITCRASQSISDYLHWYQQ




KPGKAPKLLIKYASQSISGVPSRFSGSGSGTDFTLTISSLQPEDFAT




YYCQDSHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGR




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


480
EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
CD28scfv-



RAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTVSSGGGGSG
Fc



GGGSGGGGSGGGGSQAVVTQERSLTVSPGGTVTLTCGSSTGAV




TTSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKA




ALTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLSGGGGSQV




QLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWRQAPGQGLE




WIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDDTA




VYFCTRSHYGLDWNFDVWGQGTTVTVSSGKPGSGKPGSGKPGS




GKPGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQQKP




GKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYY




CQQGQTYPYTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPGK



 


481
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQG
CD28scfv-



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
CD3scfv-Fc



TAVYFCTRSHYGLQWNFDVWGQGTTVTVSSGKPGSGKPGSGKP




GSGKPGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWYQ




QKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPEDF




ATYYCQQGQTYPYTFGGGTKVEIKSGGGGSEVQLVESGGGLVQR




GGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNY




ATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGN




FGDSYVSWFEYWGQGTLVTVSSGGGGSGGGGGGGGGGGGS




QAVVTQFPSLTVSPGGTVTLTCGSSTGAVTTSNYANWWVQQKPGK




SPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYY




CALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPS




VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV




HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL




PASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYP




SDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQ




GNVFSCSVMHEALHNHYTQKSLSLSPGK



 


482
DVQLQESGPGLVKPSQSLSLTCTVTGYSITSDYAWNWIRQFPGNK
HC; CD28



LEWMGYIRYSGSTSYNPSLKSRISITRDTSKNQFFLQLNSVTTEDT




ATYYCARDWPRPSYWYFDVWGAGTTVTVSSASTKGPSVFPLAPS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTGVVVDVS




HEDREVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GK



 


483
DIQTTQTTSSLSASLGDRVTISCRAGQDISNYLNWYQQKPDGTVKL
LC-CD28



LIYYTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGH




TLPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN




FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK




ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC



 


484
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQAPGK
CD137.39scfv-



GLEWVADIKNDGSYTNYAPSLINRFTISRDNAKNSLYLQMNSLRA
CD3scfv-



EDTAVYYCARELTGTWGQGTMVTVSBGGGGSGGGGGGGGSG
Fc



GGGSDIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAW




YQQKPGQPPKLLIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAE




DVAVYYCLQYDRYPFTFGQGTKLEIKSGGGGSEVQLVESGGGLV




QPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYN




NYATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRH




GNFGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPG




SQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG




KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADY




YCALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGP




SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVE




VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA




LPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFY




PSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ




QGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


486
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQ
CD137.b



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
scfv-



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGGGGGGGGGGGG
CD3scfv-Fc



GGSGGGGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQ




QKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFA




VYYCQDGHSFPPTFGGGTKVEIKSGGGGSEVQLVESGGGLVQPG




GSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWYGRIRSKYNNYA




TYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNF




GDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQA




VVTQFPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSP




RGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA




LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVF




LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN




AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAS




IEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIA




VEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNV




FSCSVMHEALHNHYTQKSLSLSPGK



 


487
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
CD137.U-



LEWIGEINHGGYVTYNPSLESRVTISVDTSKNQFSLKLSSVTAADTA
Fc-CD3scfv



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSASTKGPSVFPLARS




SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVERKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRK




KLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSD




GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP




GKSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMN




VWRQAPGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKN




TLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTV




SSGGGGGGGGSGGGSGGGGSQAVVTQERSLTVSPGGTVTLT




CGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFS




GSLLGGKAALTISGAQPEDEADYYCALWYSNHWWFGGGTKLTVL



 


488
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTUMISRTPEVTC
Fc-



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
CD137.Bscfv



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTL




PPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPP




VLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQES




LSLSPGKSGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFSSY




WMHWWVRQAPGQRLEWMGEINPGNGHTNYSQKFQGRVTITVDKS




ASTAYMELSSLRSEDTAVYYCARSFTTARAFAYWGQGTLVTVSSG




KPGSGKPGSGKPGSGKPGSDIVMTQSPPTLSLSPGERVTLSCRAS




QSISDYLHWYQQKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFT




LTISSLEPEDFAVYYCQDGHSFPPTFGGGTKVEIK



 


489
EPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTG
Fc-



VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
CD137.Bscfv-



LTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTL
CD137.Bscfv



PPSRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPP




VLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQES




LSLSPGKSGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFSSY




WMHWVRQAPGQRLEWMGEINPGNGHTNYSQKFQGRVTITVDKS




ASTAYMELSSLRSEDTAVYYCARSFTTARAFAYWGQGTLVTVSSG




KPGSGKPGSGKPGSGKPGSDIVMTQSPPTLSLSPGERVTLSCRAS




QSISDYLHWYQQKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFT




LTISSLEREDFAVYYCQDGHSFPPTFGGGTKVEIKSGGGSQVQLV




QSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQRLEW




MGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRSEDTA




VYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGK




PGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQKPGQ




SPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ




DGHSFPPTFGGGTKVEIK



 


490
EVQLVESGGGLVQPGGSLRLSCAASGFTESTYAMNWVRQAPGK
CD3scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
CD137.39scfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTVSSGGGGSG




GGGSGGGGGGGGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAV




TTSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLQGKA




ALTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLSGGGGSEV




QLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQARGKGL




EWVADIKNDGSYTNYAPSLINRFTISRDNAKNSLYLQMNSLRAED




TAVYYCARELTGTWGQGTMVTVSSGKPGSGKPGSGKPGSGKPG




SDIVMTQSPDSLAVSLGERATINCKSSQSLLSSGNQKNYLAWYQQ




KPGQPPKLLIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAEDVA




VYYCLQYDRYPFTFGQGTKLEIKEPKSSDKTHTCPPCPAPEFEGG




PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV




EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK




ALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGF




YPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRW




QQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


491
EVQLVESGGGLVQRGGSLRLSCAASGFTFSTYAMNWVRQAPGK
CD3scfv-



GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSL
CD137.Bscfv-Fc



RAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTVSSGGGGSG




GGGSGGGGSGGGGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAV




TTSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKA




ALTISGAQREDEADYYCALWYSNHWVFGGGTKLTVLSGGGGSQV




QLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQRL




EWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRSED




TAVYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKPGS




GKPGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQKR




GQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAVYY




CQDGHSFPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSV




FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVEVH




NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA




SIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDI




AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN




VFSCSVMHEALHNHYTQKSLSLSPGK



 


492
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNFGITWVRQAPGQG
CLDN182scfv-



LEWMGBYPSSGNTFYNEKFKGRVTMTTDTSTSTAYMELRSLRSD
CD3scfv-Fc



DTAVYYCARGGGPLRSRYFDYWGQGTLVTVSSGGGGSGGGGSG




GGGSGGGGSDIVMTQSPDSLAVSLGERATINCRSSQSLFSSGNQ




KNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLT




ISSLQAEDVAVYYCQNDYYYPLTFGGGTKVEIKSGGGGSEVQLVE




SGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVG




RIRSKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAV




YYCVRHGNFGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKP




GSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANW




VQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQR




EDEADYYCALWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAP




EFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW




YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC




KVSNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTC




LVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTV




DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


493
QVQLVQSGAEVKKPGASVKVSCKASGYTFTNSGMNWVRQAPGQ
CLDN182scfv-



GLEWMGWINTNTGEPTFAEEFRGRVTMTRDTSISTAYMELSRLRS
CD3scfv-Fc



DDTAVYYCARYYYGNSFAYWGQGTLVTVSSGGGGGGGGGGGG




GGSGGGGSDIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKN




YLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTIS




SLQAEDVAVYYCQNNYFYPLTFGGGTKVEIKSGGGGSEVQLVESG




GGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIR




SKYNNYATYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYY




CVRHGNFGDSYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGS




GKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ




QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED




EADYYCALWYSNHWVFGGGTKLTVLERKSSDKTHTCPPCPAPEF




EGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV




DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV




SNKALPASIEKTISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLV




KGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDK




SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK



 


494
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQSPEKG
CD137.Ufab-



LEWIGEINHGGYVTYNPSLESRVTISVQTSKNQFSLKLSSVTAADTA
Fc--



VYYCARDYGPGNYDWYFDLWGRGTLVTVSSASTKGPSVFPLARS
CD3scfv-



SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS
DLL3scfv



SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTGVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCRK




KLTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSD




GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP




GKSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMN




WVRQAPGKGLEWVGRIRSKYNNYATYYADSVKGRATISRDDSKN




TLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTV




SSGGGGSGGGGSGGGGGGGGGSQAVVTQEPSLTVSPGGTVTLT




CGSSTGAVTTSNYANWWQQKPGKSPRGLIGGTNKRAPGVPARFS




GSLLGGKAALTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLS




GGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVR




QAPGQGLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMEL




SSLRSEDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGS




GGGGSGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAW




YQQKPGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPE




DFATYYCQQYSTYPYTFGQGTKLEIK



 


495
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQ
CD137.Bfab-



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
Fc-



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSASTKGPSVFPLAPS
CDSscfv



SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS




SGLYSLSSVVTVPSSSIGTQTYICNVNHKPSNTKVQKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKENWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLDSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GKSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTESTYAMN




WWRQAPGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKN




TLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTV




SSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSLTVSPGGTVTLT




CGSSTGAVTTSNYANWVQQKRGKSPRGLIGGTNKRAPGVPARFS




GSLLGGKAALTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVL



 


496
QVQLVQSGAEVKKPGASVKVSCKASGYTESSYWMHWVRQAPGQ
CD137.Bfab-



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
Fc--



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSASTKGPSVFPLAPS
CD3scfv-



SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS
DLL3scfv



SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD




KTHTCPPCPAPEFEGGPSVFLAPPKPKDTLMISRTPEVTCVVVDVS




HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ




DWINGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSRD




ELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVLQSD




GSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLSLSP




GKSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMN




WRQAPGKGLEWVGRIRSKYNNYATYYADSVKGRATISRDDSKN




TLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFEYWGQGTLVTV




SSGGGGSGGGGSGGSGGGGSQAVVTQEPSLTVSPGGTVTLT




CGSSTGAVTTSNYANWVQQKFGKSPRGLIGGTNKRAPGVPARFS




GSLLGGKAALTISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLS




GGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVR




QAPGQGLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMEL




SSLRSEDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGS




GGGGSGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAW




YQQKPGKAPKLLIYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPE




DFATYYCQQYSTYPYTFGQGTKLEIK



 


497
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3scfv-



GLEWMGVINPYNDITIYNQKFQGRVTMTVQRSTSTVYMELSSLRS
CD3scfv-



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSGGGGGGGG
Fc-



SGGGGSDIQLTQSPSFLSASVGDRVTITCKASQNVGIAVAWYQQK
CD137.Bscfv



PGKAPKLUYAASNRYTGVPSRFSGSGSGTEFTLTISSLQPEDFAT




YYCQQYSTYPYTFGQGTKLEIKSGGGSEVQLVESGGGLVQPGGS




LRLSCAASGFTFSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATY




YADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGD




SYVSWFEYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVV




TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRG




LIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW




YSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPEFEGGPSVFLAP




PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT




KPREEQYNSTYRVVSVLIVLHQDWLNGKEYKCKVSNKALPASIEK




TISKAKGQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVE




WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFS




CSVMNEALNNHYTQKSLSLSPGKSGGGSQVQLVQSGAEVKKPGA




SVKVSCKASGYTFSSYWMHWVRQAPGQRLEWMGEINPGNGHTN




YSQKFQGRVTITVDKSASTAYMELSSLRSEDTAVYYCARSFTTARA




FAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSDIVMTQSPPT




LSLSPGERVTLSCRASQSISDYLHWYQQKPGQSPRLLIKYASQSIS




GIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQDGHSFPPTFGGGT




KVEIK



 


498
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWVRQAPGQ
DLL3Fab-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
Fc-



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSASTKGPSVFP
CD137.Bscfv



LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA




VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK




SCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVV




DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV




LHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPP




SRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVL




DSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLS




LSPGKSGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFSSYW




MHWWRQAPGQRLEWMIGEINPGNGHTNYSQKFQGRVTITVDKSÅ




STAYMELSSLRSEDTAVYYCARSFTTARAFAYWGQGTLVTVSSGK




PGSGKPGSGKFGSGKPGSDIVMTQSPPTLSLSPGERVTLSCRAS




QSISDYLHWYQQKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFT




LTISSLEPEDFAVYYCQDGHSFPPTFGGGTKVEIK



 


499
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYFMNWRQAPGQ
DLL3Fab-



GLEWMGVINPYNDITIYNQKFQGRVTMTVDRSTSTVYMELSSLRS
Fc-



EDTAVYYCAREGVLYDGYYEGAYWGQGTLVTVSSASTKGPSVFP
CD137.Bscfv-



LARSSKSTSGGTAALGCLVKDYFREPVTVSWNSGALTSGVHTFRA
CD137.Bscfv



VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK




SCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVV




DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV




LHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPP




SRDELTKNQVSLTCLVEGFYPSDIAVEWESNGQPENNYKTTPPVL




DSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQESLS




LSPGKSGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFSSYW




MHWVRQAPGQRLEWMGEINPGNGHTNYSQKFQGRVTITVDKSA




STAYMELSSLRSEDTAVYYCARSFTTARAFAYWGQGTLVTVSSGK




PGSGKPGSGKPGSGKPGSDIVMTQSPPTLSLSPGERVTLSCRAS




QSISDYLHWYQQKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFT




LTISSLEPEDFAVYYCQDGHSFPPTFGGGTKVEIKSGGGSQVQLV




QSGAEVKKPGASVKVSCKASGYTFSSYWMHWRQAPGQRLEW




MGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRSEDTA




VYYCARSFTTARAFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGK




PGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQQKPGQ




SPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQ




DGHSFPPTFGGGTKVEIK



 


500
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYINWVRQAPGQG
CD28(C50S)



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
scfv-



TAVYFCTRSHYGLDWNFDVWGQGTTVTVSSGGGGSGGGGSGG
CD137.Bscfv-



GGSGGGGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWY
Fc



QQKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPED




FATYYCQQGQTYPYTFGGGTKVEIKSGGGGSQVQLVQSGAEVKK




PGASVKVSCKASGYTFSSYWMHWVRQAPGQRLEWMGEINPGNG




HTNYSQKFQGRVTITVQKSASTAYMELSSLRSEDTAVYYCARSFTT




ARAFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSDIVMTQS




PPTLSLSPGERVTLSCRASQSISDYLHWYQQKPGQSPRLLIKYASQ




SISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQDGHSFPPTFG




GGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMI




SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN




STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQP




REPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPE




NNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL




HNHYTQKSLSLSPGK



 


501
QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYINWVRQAPGQG
CD28(C50S)



LEWIGSIYPGNVNTNYNEKFKDRATLTVDTSISTAYMELSRLRSDD
scfv-



TAVYFCTRSHYGLDWNFDVWGQGTTVTVSSGGGGSGGGGSGG
CD137.39scfv-



GGSGGGGSDIQMTQSPSSLSASVGDRVTITCHASQNIYVWLNWY
Fc



QQKPGKAPKLLIYKASNLHTGVPSRFSGSGSGTDFTLTISSLQPED




FATYYCQQGQTYPYTFGGGTKVEIKSGGGGSEVQLVESGGGLVQ




PGGSLRLSCAASGFTFSDYWMSWVRQAPGKGLEWVADIKNDGS




YTNYAPSLTNRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARELTG




TWGQGTMVTVSSGKPGSGKPGSGKPGSGKPGSDIVMTQSPDSL




AVSLGERATINCKSSQSLLSSGNQKNYLAWYQQKRGQPPKLLIYY




ASTRQSGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCLQYDRYP




FTFGQGTKLEIKEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKD




TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE




QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAK




GQPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNG




QPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH




EALHNHYTQKSLSLSPGK



 


502
QVQLVQSGAEVKKPGASVKVSCKASGYTFSSYWMHWVRQAPGQ
CD137.Bscfv-



RLEWMGEINPGNGHTNYSQKFQGRVTITVDKSASTAYMELSSLRS
CD28(C50S)



EDTAVYYCARSFTTARAFAYWGQGTLVTVSSGGGGSGGGGSGG
scfv-Fc



GGSGGGGSDIVMTQSPPTLSLSPGERVTLSCRASQSISDYLHWYQ




QKPGQSPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSLEPEDFA




VYYCQDGHSFPPTFGGGTKVEIKSGGGGSQVQLVQSGAEVKKPG




ASVKVSCKASGYTFTSYYIHWVRQAPGQGLEWIGSIYPGNVNTNY




NEKFKDRATLTVDTSISTAYMELSRLRSDDTAVYFCTRSHYGLDW




NFDVWGQGTTVTVSSGKPGSGKPGSGKPGSGKPGSDIQMTQSP




SSLSASVGDRVTITCHASQNIYVWLNWYQQKPGKAPKLUYKASNL




HTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGQTYPYTFG




GGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMI




SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN




STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQP




REPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQPE




NNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL




HNHYTQKSLSLSPGK



 


503
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYWMSWVRQAPGK
CD137.39scfv-



GLEWVADIKNDGSYTNYAPSLTNRFTISRDNAKNSLYLQMNSLRA
CD28(C50S)



EDTAVYYCARELTGTWGQGTMVTVSSGGGGSGGGGSGGGGSG
scfv-Fc



GGGSDIVMTQSPDSLAVSLGERATINCKSSQSLUSSGNQKNYLAW




YQQKPGQPPKLLIYYASTRQSGVPDRFSGSGSGTDFTLTISSLQAE




DVAVYYCLQYDRYPFTFGQGTKLEIKSGGGGSQVQLVQSGAEVK




KPGASVKVSCKASGYTFTSYYINWVRQAPGQGLEWIGSIYPGNVN




TNYNEKFKDRATLTVDTSISTAYMELSRLRSDDTAVYFCTRSHYGL




DWNFDVWGQGTTVTVSSGKPGSGKPGSGKPGSGKFGSDIQMTQ




SPSSLSASVGDRVTITCHASQNIYVWLNWYQQKPGKAPKLLIYKAS




NLHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGQTYPYT




FGGGTKVEIKEPKSSDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTL




MISRTPEVTGVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ




YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKG




QPREPQVYTLPPCRKKLTKNQVSLTCLVKGFYPSDIAVEWESNGQ




PENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE




ALHNHYTQKSLSLSPGK



 


504
GGGGS
linker


 


505
GKPGS
linker


 


506
GEPGS
linker


 


507
GGPGS
linker