Antibodies that bind to CTLA-4 and their uses

Abstract

Disclosed are isolated monoclonal antibodies comprising a CD152 binding domain, wherein the antibody specifically binds to human CD152. Also provided are methods of making and using the antibodies to treat diseases including cancer and autoimmune diseases.

Description

[0001] This application is a divisional application based on a patent application with an application date of December 19, 2018, a priority date of December 20, 2017, an application number of 201880083036.7, and an invention name of “Antibodies binding to CTLA-4 and their uses”. Field of the Invention

[0002] The present invention relates to monoclonal anti-CTLA-4 antibodies, nucleic acids encoding the antibodies, expression vectors and recombinant cells containing the nucleic acids, and pharmaceutical compositions containing the antibodies. Also provided are methods for preparing the antibodies and methods for using the antibodies to treat diseases including cancer and autoimmune diseases. Background of the Invention

[0003] Cancer immunotherapy is the latest breakthrough in cancer treatment, which uses the patient's own immune system to attack tumor cells. Promoting a strong CD8 T cell-dependent cytotoxic response in the tumor microenvironment is very important for generating an effective anti-tumor immune response. However, tumors tend to evade immune surveillance by utilizing T cell suppression mechanisms. Depletion of tumor-infiltrating lymphocytes (TILs) leads to anergy of cytotoxic T cells and escape of tumor cells (Wherry and Kurachi, 2015, Nat Rev Immunol., 2015, 15: 486-499; Dyck and Mills, 2017, Eur. J. Immunol., 47(5): 765-779).

[0004] Inhibitors of immune checkpoint proteins have the potential to treat a variety of tumors, such as metastatic melanoma, lung cancer, breast cancer and renal cell carcinoma. CTLA-4 (CD152) is such an inhibitory checkpoint molecule on the surface of T cells. It was originally identified by differential screening of mouse cytolytic T cell cDNA libraries (Brunet et al., 1987, Nature, 328: 267-270). It has been suggested that CTLA-4 can act as a negative regulator of T cell activation (Walunas et al., 1994, Immunity, 1: 405-413). CTLA-4 is constitutively expressed on the surface of regulatory T cells, but the number is relatively low. It is upregulated after T cell activation. After activation, CTLA-4 interacts with CD80 (B7.1) and CD86 (B7.2), which are also ligands for CD28, with much higher binding affinity than CD28 (van der Merwe et al., 1997, J Exp Med. 185:393-403; Alegre et al., 2001, Nat Rev Immunol, 1:220-228). CD28 signaling promotes T cell activation, while the interaction of CTLA-4 with its ligands B7.1 and B7.2 prevents further T cell activation.

[0005] CTLA-4 antagonists are of interest because blocking CTLA-4 with antagonists has been shown to be an effective therapy for tumors (U.S. Patent No. 6,984,720). Antagonists such as anti-CTLA-4 mAbs inhibit this surface receptor to enhance CD4 and CD8 T cell responses and reduce the suppressive function of Treg cells. In recent years, therapies based on CTLA-4 antagonists have made rapid progress. Ipilimumab Ipilimumab is a humanized antibody that blocks the action of CTLA-4, enhancing T cell responses against tumor cells. Ipilimumab was the first drug to show improvement in overall survival in patients with metastatic melanoma in a randomized, controlled Phase 3 clinical trial. It has a manageable safety profile at a dose of 3 mg / kg as monotherapy in patients previously treated with other therapies and at a dose of 10 mg / kg in combination with dacarbazine in treatment-naive patients. In addition to malignant melanoma, ipilimumab is also being developed for the treatment of prostate cancer and non-small cell lung cancer.

[0006] However, despite the above progress, there is still a desire for CTLA-4 antagonists with improved affinity, specificity, and developability. In addition, there is a need for more effective therapies involving anti-CTLA-4 antibodies that effectively inhibit CTLA-4 signaling activity in humans while causing minimal adverse side effects. SUMMARY OF THE INVENTION

[0007] Disclosed are isolated monoclonal antibodies comprising a CD152 binding domain, wherein the CD152 binding domain comprises an immunoglobulin heavy chain variable region comprising CDR1, CDR2, and CDR3, wherein CDR1, CDR2, and CDR3 each comprise an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identity to: (1) SEQ ID NOs: 4, 5, and 6; (2) SEQ ID NOs: 10, 11, and 12; (3) SEQ ID NOs: 16, 17, and 18; (4) SEQ ID NOs: 22, 23, and 24; (5) SEQ ID NOs: 28, 29, and 30; (6) SEQ ID NOs: 34, 35, and 36; (7) SEQ ID NOs: 40, 41, and 42; (8) SEQ ID NOs: 46, 47, and 48; (9) SEQ ID NOs: 51, 52, and 53; NO:52, 53 and 54; (10) SEQ ID NO:58, 59 and 60; (11) SEQ ID NO:64, 65 and 66; (12) SEQ ID NO:70, 71 and 72; (13) SEQ ID NO:76, 77 and 78; (14) SEQ ID NO:82, 83 and 84; (15) SEQ ID NO:88, 89 and 90; (16) SEQ ID NO:94, 95 and 96; (17) SEQ ID NO:100, 101 and 102; (18) SEQ ID NO:106, 107 and 108; (19) SEQ ID NO:112, 113 and 114; (20) SEQ ID NO:118, 119 and 120; (21) SEQ ID NO:124, 125 and 126; (22) SEQ ID NO:130, 131 and 132; (23) SEQ ID NO:136, 137 and 138; (24) SEQ ID NO:142, 143 and 144; (25) SEQ ID NO:148, 149 and 150; (26) SEQ ID NO:154, 155 and 156; (27) SEQ ID NO:160, 161 and 162; (28) SEQ ID NO:166, 167 and 168; (29) SEQ ID NO:172, 173 and 174; (30) SEQ ID NO:178, 179 and 180; (31) SEQ ID NO:184, 185 and 186; or (32) SEQ ID NO:190, 191 and 192.

[0008] In some cases, the antibody is a heavy chain-only antibody. In some cases, the antibody does not comprise an immunoglobulin light chain. In some cases, the antibody comprises one immunoglobulin heavy chain. In some cases, the antibody comprises two immunoglobulin heavy chains. In some cases, the antibody consists of two immunoglobulin heavy chains. In some cases, at least one of the two immunoglobulin heavy chains comprises an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 2, 8, 14, 20, 26, 32, 38, 44, 50, 56, 62, 68, 74, 80, 86, 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182 or 188.

[0009] In some cases, the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, 45, 51, 57, 63, 69, 75, 81, 87, 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183, or 189.

[0010] In some cases, the antibody specifically binds to human CD152. In some cases, the antibody binds to human CD152 with high affinity. In some cases, the antibody binds to human CD152 with a higher affinity than an ipilimumab analog. In some cases, the antibody binds to human CD152 with at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, or at least 100-fold greater affinity than an ipilimumab analog.

[0011] In some cases, the antibody is expressed as 1.0*10 -7 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -8 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -9 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -10 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -11 M or smaller K dDissociated from human CD152. In some cases, K d 6.0*10 -11 M or less. In some cases, the antibody has a lower K than the ipilimumab analog. d Dissociate from human CD152. In some cases, the antibody dissociates with a K of at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, or at least 100-fold lower than that of the ipilimumab analog. d Dissociation from human CD152. In some cases, K was determined by surface plasmon resonance. d .

[0012] In some cases, the antibody specifically binds to monkey CD152. In some cases, the antibody does not specifically bind to monkey CD152. In some cases, the antibody blocks the binding of CD152 to CD80, CD86, or both. In some cases, the antibody promotes IL-2 secretion by immune cells. In some cases, the antibody induces T cell activation. In some cases, the antibody stimulates an anti-tumor immune response by immune cells. In some cases, the antibody is human, humanized, or chimeric.

[0013] In another aspect, disclosed herein are isolated monoclonal heavy chain-only antibodies comprising a CD152 binding domain, wherein the antibody specifically binds to human CD152. In some cases, the antibody does not comprise an immunoglobulin light chain. In some cases, the antibody comprises one immunoglobulin heavy chain. In some cases, the antibody comprises two immunoglobulin heavy chains. In some cases, the antibody consists of two immunoglobulin heavy chains.

[0014] In some cases, the antibody binds to human CD152 with high affinity. In some cases, the antibody binds to human CD152 with a higher affinity than an ipilimumab analog. In some cases, the antibody binds to human CD152 with at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, or at least 100-fold greater affinity than an ipilimumab analog.

[0015] In some cases, the antibody is expressed as 1.0*10 -7 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -8 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -9 M or smaller K d Dissociation from human CD152. In some cases, the antibody was 1.0*10 -10 M or smaller K dDissociation from human CD152. In some cases, the antibody was 1.0*10 -11 M or smaller K d Dissociated from human CD152. In some cases, K d 6.0*10 -11 M or less. In some cases, the antibody has a lower K than the ipilimumab analog. d Dissociate from human CD152. In some cases, the antibody dissociates with a K of at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, or at least 100-fold lower than that of the ipilimumab analog. d Dissociation from human CD152. In some cases, K was determined by surface plasmon resonance. d .

[0016] In some cases, the antibody specifically binds to monkey CD 152. In some cases, the antibody does not specifically bind to monkey CD 152. In some instances, the CD152 binding domain comprises an immunoglobulin heavy chain variable region comprising CDR1, CDR2, and CDR3, wherein CDR1, CDR2, and CDR3 each comprise an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to: (1) SEQ ID NOs: 4, 5, and 6; (2) SEQ ID NOs: 10, 11, and 12; (3) SEQ ID NOs: 16, 17, and 18; (4) SEQ ID NOs: 22, 23, and 24; (5) SEQ ID NOs: 28, 29, and 30; (6) SEQ ID NOs: 34, 35, and 36; (7) SEQ ID NOs: 40, 41, and 42; (8) SEQ ID NOs: 46, 47, and 48; (9) SEQ ID NOs: 52, 53, and 54; (10) SEQ ID NOs: NO:58, 59 and 60; (11) SEQ ID NO:64, 65 and 66; (12) SEQ ID NO:70, 71 and 72; (13) SEQ ID NO:76, 77 and 78; (14) SEQ ID NO:82, 83 and 84; (15) SEQ ID NO:88, 89 and 90; (16) SEQ ID NO:94, 95 and 96; (17) SEQ ID NO:100, 101 and 102; (18) SEQ ID NO:106, 107 and 108; (19) SEQ ID NO:112, 113 and 114; (20) SEQ ID NO:118, 119 and 120; (21) SEQ ID NO:124, 125 and 126; (22) SEQ ID NO:130, 131 and 132; (23) SEQ ID NO: NO:136, 137 and 138; (24) SEQ ID NO:142, 143 and 144; (25) SEQ ID NO:148, 149 and 150; (26) SEQ ID NO:154, 155 and 156; (27) SEQ ID NO:160, 161 and 162; (28) SEQ ID NO:166, 167 and 168; (29) SEQ ID NO:172, 173 and 174; (30) SEQ ID NO:178, 179 and 180; (31) SEQ ID NO:184, 185 and 186; or (32) SEQ ID NO:190, 191 and 192.

[0017] In some instances, the CD152 binding domain comprises at least one immunoglobulin heavy chain, wherein the immunoglobulin heavy chain comprises an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 2, 8, 14, 20, 26, 32, 38, 44, 50, 56, 62, 68, 74, 80, 86, 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, or 188. In some cases, the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, 45, 51, 57, 63, 69, 75, 81, 87, 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183, or 189.

[0018] In some cases, the antibodies block the binding of CD152 to CD80, CD86, or both. In some cases, the antibodies promote the secretion of IL-2 by immune cells. In some cases, the antibodies induce T cell activation. In some cases, the antibodies stimulate an anti-tumor immune response by immune cells. In some cases, the antibodies are human, humanized, or chimeric antibodies.

[0019] On the other hand, disclosed herein is a pharmaceutical composition comprising any antibody disclosed herein and a pharmaceutically acceptable excipient thereof. In some cases, the pharmaceutically acceptable excipient is selected from the group consisting of a carrier, a surfactant, a thickener or emulsifier, a solid binder, a dispersion or suspension aid, a solubilizer, a colorant, a flavoring, a coating, a disintegrant, a lubricant, a sweetener, a preservative, an isotonic agent, and a combination thereof. In some cases, the pharmaceutical composition further comprises a second antibody, wherein the second antibody is an immunostimulatory antibody or a co-stimulatory antibody. In some cases, the immunostimulatory antibody is selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-LAG-3 antibody, an anti-TIM3 antibody, an anti-STAT3 antibody, and an anti-ROR1 antibody. In some cases, the co-stimulatory antibody is an anti-CD137 antibody or an anti-GITR antibody.

[0020] In another aspect, disclosed herein are isolated nucleic acid molecules encoding any of the antibodies disclosed herein. In some cases, the nucleic acid molecule comprises a nucleotide sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 163, 169, 175, 181 or 187. In some cases, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 163, 169, 175, 181, or 187.

[0021] In another aspect, disclosed herein are expression vectors comprising a nucleic acid segment encoding any of the antibodies disclosed herein, wherein the nucleic acid segment is operably linked to a regulatory sequence suitable for expressing the nucleic acid segment in a host cell. In some cases, the nucleic acid segment comprises a nucleotide sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 163, 169, 175, 181, or 187. In some cases, the nucleic acid segment comprises the nucleotide sequence shown in SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 163, 169, 175, 181, or 187.

[0022] In another aspect, disclosed herein is a host cell comprising any of the expression vectors disclosed herein.

[0023] In another aspect, disclosed herein are methods for producing CD152-binding monoclonal antibodies, comprising culturing a host cell comprising any expression vector under conditions that express a nucleic acid segment, thereby producing the CD152-binding monoclonal antibody. In some cases, the host cell is a CHO, HEK293, or COS host cell line. In some cases, the CHO host cell line is a CHO-K1 cell line. In some cases, the method further comprises recovering the CD152-binding monoclonal antibody.

[0024] In another aspect, disclosed herein is a method of inducing antibody-dependent cell-mediated cytotoxicity (ADCC) against cells expressing a tumor-associated antigen, the method comprising contacting a T cell with any of the antibodies disclosed herein, wherein the contacting is performed under conditions that induce ADCC against cells expressing the tumor-associated antigen.

[0025] On the other hand, disclosed herein is a method for treating a condition in a subject, the method comprising administering to the subject a therapeutically effective amount of any antibody disclosed herein or any pharmaceutical composition disclosed herein. In some cases, the condition is cancer. In some cases, the cancer is selected from the group consisting of leukemia, lymphoma, CLL, small lymphocytic lymphoma, marginal cell B-cell lymphoma, Burkitt's lymphoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, myeloma, gastric cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, and head and neck cancer. In some cases, the method further comprises a therapeutic agent. In some cases, the therapeutic agent is an anticancer drug. In some cases, the therapeutic agent is ipilimumab or its biosimilar product. In some cases, the condition is an autoimmune disease.

[0026] On the other hand, disclosed herein is the use of any pharmaceutical composition disclosed herein in the preparation of a medicament for treating a disease. In some cases, the disease is cancer. In some cases, the cancer is selected from the group consisting of leukemia, lymphoma, CLL, small lymphocytic lymphoma, marginal cell B-cell lymphoma, Burkitt's lymphoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, myeloma, gastric cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, and head and neck cancer. In some cases, the disease is an autoimmune disease.

[0027] In another aspect, the present invention relates to an isolated monoclonal antibody, or antigen-binding portion thereof, having a heavy chain variable region comprising a CDR1 region, a CDR2 region, and a CDR3 region, wherein the CDR1 region, the CDR2 region, and the CDR3 region, respectively, comprise an amino acid sequence that is at least 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to: (1) SEQ ID NOs: 4, 5, and 6; (2) SEQ ID NOs: 10, 11, and 12; (3) SEQ ID NOs: 16, 17, and 18; (4) SEQ ID NOs: 22, 23, and 24; (5) SEQ ID NOs: 28, 29, and 30; (6) SEQ ID NOs: 34, 35, and 36; (7) SEQ ID NOs: 40, 41, and 42; (8) SEQ ID NOs: 46, 47, and 48; (9) SEQ ID NOs: 52, 53, and 54; (10) SEQ ID NOs: NO:58, 59 and 60; (11) SEQ ID NO:64, 65 and 66; (12) SEQ ID NO:70, 71 and 72; (13) SEQ ID NO:76, 77 and 78; (14) SEQ ID NO:82, 83 and 84; (15) SEQ ID NO:88, 89 and 90; (16) SEQ ID NO:94, 95 and 96; (17) SEQ ID NO:100, 101 and 102; (18) SEQ ID NO:106, 107 and 108; (19) SEQ ID NO:112, 113 and 114; (20) SEQ ID NO:118, 119 and 120; (21) SEQ ID NO:124, 125 and 126; (22) SEQ ID NO:130, 131 and 132; (23) SEQ ID NO: NO:136, 137 and 138; (24) SEQ ID NO:142, 143 and 144; (25) SEQ ID NO:148, 149 and 150; (26) SEQ ID NO:154, 155 and 156; (27) SEQ ID NO:160, 161 and 162; (28) SEQ ID NO:166, 167 and 168; (29) SEQ ID NO:172, 173 and 174; (30) SEQ ID NO:178, 179 and 180; (31) SEQ ID NO:184, 185 and 186; or (32) SEQ ID NO:190, 191 and 192; wherein the antibody or antigen-binding fragment thereof binds to CTLA-4.

[0028] In another aspect, the isolated monoclonal antibody of the invention, or antigen binding portion thereof, comprises a heavy chain variable region comprising an amino acid sequence that is at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, 45, 51, 57, 63, 69, 75, 81, 87, 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183 or 189.

[0029] In one embodiment, an antibody of the invention comprises a heavy chain comprising an amino acid sequence that is at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO: 2, 8, 14, 20, 26, 32, 38, 44, 50, 56, 62, 68, 74, 80, 86, 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182 or 188, which may be composed of an amino acid sequence that is at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO: 2, 8, 14, 20, 26, 32, 38, 44, 50, 56, 62, 68, 74, 80, 86, 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170 NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 157, 163, 169, 175, 181 or 187 is encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% or 100% identity.

[0030] In some embodiments, an antibody of the invention consists essentially of or consists of two heavy chains as described above.

[0031] In another aspect of the present invention, antibody or its antigen binding portion thereof is a part of an immunoconjugate, which includes a therapeutic agent connected to an antibody, such as a cytotoxin or a radioisotope. On the other hand, antibody is a part of a bispecific molecule, which includes a second functional portion (e.g., a second antibody) having different binding specificities from the antibody or its antigen binding portion thereof. On the other hand, the antibody of the present invention or its antigen binding portion thereof can be prepared as a part of a chimeric antigen receptor (CAR) or an engineered T cell receptor. The antibody of the present invention or its antigen binding portion thereof can also be encoded by an oncolytic virus or used in combination with an oncolytic virus.

[0032] Also provided are pharmaceutical compositions comprising an antibody, or antigen-binding portion thereof, immunoconjugate, bispecific molecule, chimeric antigen receptor, engineered T cell receptor, or oncolytic virus of the invention, optionally formulated in a pharmaceutically acceptable carrier.

[0033] The pharmaceutical composition may further comprise other anticancer agents. The pharmaceutical composition may further comprise at least one additional immunostimulatory antibody selected from the group consisting of anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-LAG-3 antibodies, anti-TIM3 antibodies, anti-STAT3 antibodies, and anti-ROR1 antibodies, and / or a co-stimulatory antibody, which may be an anti-CD137 antibody or an anti-GITR antibody.

[0034] Also provided are nucleic acid molecules encoding the antibodies of the invention or antigen-binding portions thereof (e.g., variable regions and / or CDRs), as well as expression vectors comprising the nucleic acids and host cells comprising the expression vectors. Also provided are methods for preparing anti-CTLA-4 antibodies using host cells comprising the expression vectors, and the methods comprise the steps of: (i) expressing the antibody in the host cell, and (ii) isolating the antibody from the host cell.

[0035] In another embodiment, the present invention provides a method for inhibiting the growth of tumor cells in a subject, comprising administering to the subject a therapeutically effective amount of an antibody of the present invention or its antigen-binding portion thereof. The tumor can be a solid tumor or a non-solid tumor, selected from the group consisting of leukemia, lymphoma, CLL, small lymphocytic lymphoma, marginal cell B-cell lymphoma, Burkitt's lymphoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, myeloma, gastric cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, and head and neck cancer. Preferably, the tumor is melanoma, prostate cancer, or non-small cell lung cancer. In another embodiment, the present invention provides a method for treating an autoimmune disease in a subject, comprising administering to the subject a therapeutically effective amount of an antibody of the present invention or its antigen-binding portion thereof. In another embodiment, the present invention provides a method for treating a viral infection in a subject, comprising administering to the subject a therapeutically effective amount of an antibody of the present invention or its antigen-binding portion thereof. In another embodiment, the method comprises administering a pharmaceutical composition, bispecific, or immunoconjugate of the present invention.

[0036] In another aspect, the invention provides anti-CTLA-4 antibodies and compositions of the invention for use in the aforementioned methods, or for the preparation of a medicament for use in the aforementioned methods (e.g., for treatment).

[0037] Other features and advantages of the present disclosure will become apparent through the following detailed description and examples, which should not be construed as limiting. The contents of all references, Genbank entries, patents, and published patent applications cited in this application are expressly incorporated herein by reference.

[0038] The present invention includes the following embodiments:

[0039] Embodiment 1. An isolated monoclonal antibody comprising a CD152 binding domain, wherein the CD152 binding domain comprises an immunoglobulin heavy chain variable region comprising CDR1, CDR2, and CDR3, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence having at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identity to:

[0040] (1) SEQ ID NO: 4, 5 and 6;

[0041] (2) SEQ ID NOs: 10, 11, and 12;

[0042] (3) SEQ ID NO: 16, 17 and 18;

[0043] (4) SEQ ID NO: 22, 23 and 24;

[0044] (5) SEQ ID NO: 28, 29 and 30;

[0045] (6) SEQ ID NO: 34, 35 and 36;

[0046] (7) SEQ ID NO: 40, 41 and 42;

[0047] (8) SEQ ID NO: 46, 47 and 48;

[0048] (9) SEQ ID NO: 52, 53 and 54;

[0049] (10) SEQ ID NO: 58, 59 and 60;

[0050] (11) SEQ ID NO: 64, 65 and 66;

[0051] (12) SEQ ID NO: 70, 71 and 72;

[0052] (13) SEQ ID NO: 76, 77 and 78;

[0053] (14) SEQ ID NO: 82, 83 and 84;

[0054] (15) SEQ ID NO: 88, 89 and 90;

[0055] (16) SEQ ID NO:94, 95 and 96;

[0056] (17) SEQ ID NO: 100, 101 and 102;

[0057] (18) SEQ ID NO: 106, 107 and 108;

[0058] (19) SEQ ID NO: 112, 113 and 114;

[0059] (20) SEQ ID NO: 118, 119 and 120;

[0060] (21) SEQ ID NO: 124, 125 and 126;

[0061] (22) SEQ ID NO: 130, 131 and 132;

[0062] (23) SEQ ID NO: 136, 137 and 138;

[0063] (24) SEQ ID NO: 142, 143 and 144;

[0064] (25) SEQ ID NO: 148, 149 and 150;

[0065] (26) SEQ ID NO: 154, 155 and 156;

[0066] (27) SEQ ID NO: 160, 161 and 162;

[0067] (28) SEQ ID NO: 166, 167 and 168;

[0068] (29) SEQ ID NO: 172, 173 and 174;

[0069] (30) SEQ ID NO: 178, 179 and 180;

[0070] (31) SEQ ID NO: 184, 185 and 186; or

[0071] (32)SEQ ID NO:190, 191 and 192.

[0072] Embodiment 2. The isolated monoclonal antibody of embodiment 1, wherein the antibody is a heavy chain-only antibody.

[0073] Embodiment 3. The isolated monoclonal antibody of embodiment 1 or 2, wherein said antibody comprises two immunoglobulin heavy chains.

[0074] Embodiment 4. The isolated monoclonal antibody of any one of embodiments 1 to 3, wherein said antibody consists of two immunoglobulin heavy chains.

[0075] Embodiment 5. The isolated monoclonal antibody of embodiment 3 or 4, wherein at least one of the two immunoglobulin heavy chains comprises an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 2, 8, 14, 20, 26, 32, 38, 44, 50, 56, 62, 68, 74, 80, 86, 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182 or 188.

[0076] Embodiment 6. The isolated monoclonal antibody of any one of embodiments 1 to 5, wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, 45, 51, 57, 63, 69, 75, 81, 87, 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183 or 189.

[0077] Embodiment 7. The isolated monoclonal antibody of any one of Embodiments 1 to 6, which exhibits one or a combination of the following properties: (a) binding to human CD152; (b) specific binding to monkey CD152; (c) no binding to mouse CD152; (d) blocking the binding of CD152 to CD80, CD86, or both; (e) promoting IL-2 secretion by immune cells; (f) inducing T cell activation; (g) stimulating anti-tumor immune responses of immune cells.

[0078] Embodiment 8. The isolated monoclonal antibody of any one of embodiments 1 to 7, which exhibits one or a combination of the following properties: (a) binds to human CD152 with an affinity greater than that of an ipilimumab analog; (b) binds to human CD152 with an affinity at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, or at least 100-fold greater than that of an ipilimumab analog; (c) binds to human CD152 with an affinity of 1.0*10 -9 M or smaller K d Dissociation from human CD152; (d) K compared to ipilimumab analogs d Lower K d Dissociation from human CD152; (e) K compared to ipilimumab analogs d A K that is at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, or at least 100-fold lowerd Dissociated from human CD152.

[0079] Embodiment 9. The isolated monoclonal antibody of any one of embodiments 1 to 8, wherein the antibody is expressed at a concentration of 6.0*10 -11 M or smaller K d Dissociated from human CD152.

[0080] Embodiment 10. The isolated monoclonal antibody of any one of embodiments 1 to 9, wherein the antibody is a human, humanized or chimeric antibody.

[0081] Embodiment 11. A pharmaceutical composition comprising the isolated monoclonal antibody of any one of embodiments 1 to 10 and a pharmaceutically acceptable excipient.

[0082] Embodiment 12. The pharmaceutical composition of embodiment 11, wherein the pharmaceutically acceptable excipient is selected from the group consisting of a carrier, a surfactant, a thickener or emulsifier, a solid binder, a dispersion or suspension aid, a solubilizer, a colorant, a flavoring agent, a coating, a disintegrant, a lubricant, a sweetener, a preservative, an isotonic agent, and combinations thereof.

[0083] Embodiment 13. The pharmaceutical composition of embodiment 11 or 12, further comprising a second antibody, wherein the second antibody is an immunostimulatory antibody or a co-stimulatory antibody.

[0084] Embodiment 14. The pharmaceutical composition of embodiment 13, wherein the immunostimulatory antibody is selected from the group consisting of anti-PD-1 antibody, anti-PD-L1 antibody, anti-LAG-3 antibody, anti-TIM3 antibody, anti-STAT3 antibody and anti-ROR1 antibody.

[0085] Embodiment 15. The pharmaceutical composition of embodiment 13 or 14, wherein the co-stimulatory antibody is an anti-CD137 antibody or an anti-GITR antibody.

[0086] Embodiment 16. A nucleic acid molecule encoding the isolated antibody of any one of embodiments 1 to 10.

[0087] Embodiment 17. The nucleic acid molecule of embodiment 16, wherein the nucleic acid molecule comprises a nucleotide sequence that is at least 80%, 85%, 88%, 90%, 92%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 163, 169, 175, 181 or 187.

[0088] Embodiment 18. An expression vector comprising the nucleic acid molecule of embodiment 16 or 17, wherein the nucleic acid molecule is operably linked to regulatory sequences suitable for expressing the nucleic acid segment in a host cell.

[0089] Embodiment 19. A host cell comprising the expression vector of embodiment 18.

[0090] Embodiment 20. A method of inducing antibody-dependent cell-mediated cytotoxicity (ADCC) against cells expressing a tumor-associated antigen, the method comprising contacting T cells with the isolated antibody of any one of embodiments 1 to 10, wherein the contacting is performed under conditions that induce ADCC against cells expressing the tumor-associated antigen.

[0091] Embodiment 21. Use of the isolated monoclonal antibody of any one of embodiments 1 to 10 or the pharmaceutical composition of any one of embodiments 11 to 15 in the preparation of a medicament for treating a disorder, wherein the disorder is cancer or an autoimmune disease.

[0092] Embodiment 22. The use of embodiment 21, wherein the disorder is cancer.

[0093] Embodiment 23. The use of embodiment 22, wherein the cancer is selected from the group consisting of leukemia, lymphoma, CLL, small lymphocytic lymphoma, marginal cell B-cell lymphoma, Burkitt lymphoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, myeloma, gastric cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, and head and neck cancer.

[0094] Embodiment 24. The use according to any one of embodiments 21 to 23, wherein the medicament further contains an additional therapeutic agent.

[0095] Embodiment 25. The use of embodiment 24, wherein the additional therapeutic agent is an anti-cancer agent.

[0096] Embodiment 26. The use of embodiment 24, wherein the additional therapeutic agent is ipilimumab and its biosimilar products.

[0097] Embodiment 27. The use according to embodiment 21, wherein the disorder is an autoimmune disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0098] The foregoing summary of the invention, as well as the following detailed description of the invention, will be better understood when explained in conjunction with the accompanying drawings.It should be understood that the present invention is not limited to the embodiments shown in the accompanying drawings.

[0099] In the attached figure:

[0100] Figure 1 The binding activity of the anti-CTLA-4 antibody of the present invention to human CTLA-4 expressed on 293 cells was demonstrated.

[0101] Figure 2 The binding activity of the anti-CTLA-4 antibody of the present invention to cynomolgus monkey CTLA-4 expressed on 293 cells was shown.

[0102] Figure 3 The ability of the anti-CTLA-4 antibodies of the present invention to block the interaction between CTLA-4 and B7.1 is demonstrated.

[0103] Figure 4 A and 4B show the effects of anti-CTLA-4 antibodies CL3, CL5, CL11, and CL22 (A) and CL24, CL25, and CL30 (B) on IL-2 secretion in a SEB-dependent T lymphocyte stimulation assay using PBMCs from Donor 1.

[0104] Figure 5 A and 5B show the binding ability of anti-CTLA-4 antibodies CL5, CL11, CL22, CL24, and CL25 (A) and CL5 (B) to human CTLA-4 or mouse CTLA-4.

[0105] Figure 6 A and 6B show the effects of anti-CTLA-4 antibodies CL5, CL11, and their mutants (A) and CL22, CL25, and their mutants (B) on IL-2 secretion in a SEB-dependent T lymphocyte stimulation assay using PBMCs from donor 2.

[0106] Figure 7 The binding activity of anti-CTLA-4 antibody mutants to human CTLA-4 is demonstrated.

[0107] Figure 8 The activity of anti-CTLA-4 antibody mutants in blocking the interaction between CTLA-4 and biotinylated B7.1 and B7.2 is shown.

[0108] Figure 9 Shown are the effects of anti-CTLA-4 antibody mutants on IL-2 secretion in a SEB-dependent T lymphocyte stimulation assay using PBMCs from donor 3.

[0109] Figure 10 Shown are the binding activities of the anti-CTLA-4 antibody CL5 and its mutants to human or cynomolgus monkey CTLA-4 as measured by BiaCore.

[0110] Figure 11The in vitro ADCC activity of the anti-CTLA-4 antibody CL5 and its mutants on CHO K1-CTLA-4 cells is shown.

[0111] Figure 12 Shown are the serum concentration-time profiles of the anti-CTLA-4 antibody CL5 in male C57BL / 6 mice.

[0112] Figure 13 Tumor-to-serum ratios of anti-CTLA4 HCAb concentrations are shown.

[0113] Figure 14 Figures 14A, 14B, and 14C show the antitumor activity of anti-CTLA-4 antibodies in MC38 tumor-bearing mice. (A) Tumor growth curves in different groups. (B) Kaplan-Meier survival curves of the time to endpoint. (C) Mouse body weight remained relatively constant following treatment with human anti-CTLA-4 antibodies. Data are presented as mean ± SEM, N = 9.

[0114] Figure 15 Figures A and 15B show the inhibition of tumor growth by human anti-CTLA-4 antibodies in MC38-bearing mice. (A) Tumor growth curve. (B) Mouse body weight remained relatively constant following treatment with human anti-CTLA-4 antibodies. Data are presented as mean ± SEM, N = 6. Detailed Description of the Invention

[0115] General Overview

[0116] The present disclosure provides antibodies that specifically bind to CD152. These binding molecules can specifically bind to CD152 and another target. The CD152 binding antibodies administered to patients in need thereof in a therapeutically effective amount can be used to treat certain conditions, including certain cancers. The combination of antibodies and T cells expressing CD152 induces antibody-dependent cell-mediated cytotoxicity against cells expressing tumor-associated antigens. The CD152 binding therapy disclosed herein provides a variety of advantages in treating patients, for example, effectively binding to CD152, effectively inducing antibody-dependent cell-mediated cytotoxicity and / or adverse events (such as toxicity) risk is lower. In some aspects, CD152 binding antibodies are more effectively combined with CD152 in some forms (for example, only heavy chain antibodies compared to typical full-length antibodies), resulting in higher efficacy and improved treatment of the utility of conditions related to CD152.

[0117] definition

[0118] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited herein, including but not limited to patents, patent applications, articles, books, and monographs, are expressly incorporated herein by reference in their entirety for any purpose. If one or more incorporated documents or portions of documents define a term that contradicts the definition of that term in this application, the definition appearing in this application shall prevail. However, the mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not to be considered, nor should it be considered, an admission or any form of suggestion that they constitute valid prior art or form part of the common general knowledge of any country in the world.

[0119] In this specification, unless otherwise stated, any concentration range, percentage range, ratio range or integer range should be understood to include the value of any integer within the range, and its fraction (such as one tenth and one hundredth of an integer) where appropriate. It should be understood that, unless otherwise stated, the terms "one" and "a" used herein refer to "one or more" of the listed components. The use of alternatives (such as "or") should be understood to mean one, both or any combination thereof. As used herein, the terms "comprising" and "including" are used synonymously. In addition, it should be understood that the present application discloses polypeptides comprising various combinations of components (e.g., domains or regions) and substituents as described herein, to the same extent as each polypeptide is listed separately. Therefore, the selection of specific components of a single polypeptide is within the scope of this disclosure.

[0120] As used herein, the term "about" and its grammatical equivalents relative to a reference value may include a range of values ​​plus or minus 10% from that value, such as a range of values ​​plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% from that value. For example, the number "about 10" includes numbers from 9 to 11.

[0121] As used herein, a "polypeptide" or "polypeptide chain" is a single, linear, and continuous arrangement of covalently linked amino acids. It does not include two polypeptide chains linked together in a nonlinear manner, such as via an interchain disulfide bond (e.g., half an immunoglobulin molecule in which a light chain is linked to a heavy chain via a disulfide bond). A polypeptide may have or form one or more intrachain disulfide bonds. With respect to the polypeptides described herein, references to amino acid residues corresponding to the amino acid residues designated by a SEQ ID NO include post-translational modifications of such residues.

[0122] A "protein" is a macromolecule composed of one or more polypeptide chains. Proteins may also contain non-peptide components, such as carbohydrate groups. Carbohydrate and other non-peptide substituents may be added to proteins by the cell in which they are produced and will vary with the cell type. Proteins are defined herein based on their amino acid backbone structure; substituents such as carbohydrate groups are generally not specified but may be present.

[0123] The terms "antibody," "immunoglobulin," or "Ig" are used interchangeably herein and refer to immunoglobulin molecules that recognize and specifically bind to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination thereof, through at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term "antibody" encompasses intact polyclonal antibodies, intact monoclonal antibodies, antibody fragments (such as Fab, Fab', F(ab')2, and Fμ fragments), single-chain Fμ (scFv) mutants, multispecific antibodies such as bispecific antibodies (including bispecific antibodies), chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising an antigenic determinant of an antibody, and any other modified immunoglobulin molecule comprising an antigen recognition site, as long as the antibody exhibits the desired biological activity. The term "antibody" may also refer to a Y-shaped glycoprotein with a molecular weight of approximately 150 kDa that consists of four polypeptide chains: two light (L) chains and two heavy (H) chains. There are five types of mammalian Ig heavy chain isotypes, represented by the Greek letters alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ). The heavy chain type defines the class of antibody, IgA, IgD, IgE, IgG, and IgM, respectively. The gamma and alpha classes are further divided into subclasses, such as IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2, based on differences in constant domain sequence and function. In mammals, there are two types of immunoglobulin light chains, λ and κ.

[0124] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies (i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerizations, amidations) that may be present in minor amounts). Monoclonal antibodies are highly specific for a single antigenic site. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, an advantage of monoclonal antibodies is that they are synthesized by hybridoma cultures and are not contaminated by other immunoglobulins. The modifier "monoclonal" indicates the characteristic of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies for use in accordance with the present invention can be prepared by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein, Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14(3):253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2 nded. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, NY, 1981)), recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567), phage display technology (see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34):12467-12472(2004); and Lee et al., J. Immunol. Methods 284(1-2):119-132(2004)), and techniques for producing human or human-like antibodies in animals having partial or complete human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90:2551(1993); Jakobovits et al., Nature 362:255-258(1993); Bruggemann et al., Year 2003). in Immunol. 7:33 (1993); U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al., Bio / Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859 (1994); Morrison, Nature 368:812-813 (1994); Fishwild et al., Nature Biotechnol. 14:845-851 (1996); Neuberger, Nature Biotechnol. 14:826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol.13:65-93(1995)).

[0125] As used herein, the term "heavy chain only antibody" (HCAb) refers to an antibody composed of only two heavy chains and lacking the two light chains normally present in a full-length antibody.

[0126] When used to describe the various antibodies disclosed herein, the term "isolated antibody" means an antibody that has been identified, separated and / or recovered from the cell or cell culture in which it is expressed. Contaminant components of its natural environment are substances that would generally interfere with the diagnostic or therapeutic use of the polypeptide and may include enzymes, hormones and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is purified to a purity greater than 95% or 99%, as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007). In preferred embodiments, the antibody will be purified (1) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence using a spinning cup sequencer, or (2) to homogeneity, by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.

[0127] The term "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0128] The term "light chain variable region" (also referred to as "light chain variable domain" or "VL" or V L ) and a “heavy chain variable region” (also called a “heavy chain variable domain” or “VH” or V H) refer to the variable binding regions from the light and heavy chains of an antibody, respectively. The variable binding region is composed of discrete, well-defined subregions called "complementarity determining regions" (CDRs) and "framework regions" (FRs), typically comprising FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, from the amino terminus to the carboxyl terminus. In one embodiment, the FRs are humanized. The term "CL" refers to "immunoglobulin light chain constant region" or "light chain constant region," i.e., the constant region from the antibody light chain. The term "CH" refers to "immunoglobulin heavy chain constant region" or "heavy chain constant region," which can be further divided into CH1, CH2, and CH3 (IgA, IgD, IgG) or CH1, CH2, CH3, and CH4 domains (IgE, IgM), depending on the antibody isotype. "Fab" (fragment antigen binding) is the portion of the antibody that binds to the antigen and includes the variable region and CH1 domain of the heavy chain connected to the light chain via interchain disulfide bonds.

[0129] As used herein, the term "binding domain" or "binding region" refers to a domain, region, portion or site of a binding domain of a protein, polypeptide, oligopeptide or peptide or antibody or antibody derived therefrom that has the ability to specifically recognize and bind to a target molecule such as an antigen, ligand, receptor, substrate or inhibitor (e.g., CD152). Exemplary binding domains include single-chain antibody variable regions (e.g., domain antibodies, sFv, scFv, scFab), receptor extracellular domains and ligands (e.g., cytokines, chemokines). In certain embodiments, the binding domain comprises or consists of an antigen binding site (e.g., comprising a variable heavy chain sequence and a variable light chain sequence from an antibody or three light chain complementary determining regions (CDRs) and three heavy chain CDRs, which are placed in an alternative framework region (FR) (e.g., a human FR optionally comprising one or more amino acid substitutions). A variety of assays are known for identifying binding domains of the present disclosure that specifically bind to a particular target, including Western blotting, ELISA, phage display libraries and Interaction Analysis. As used herein, a "CD152 binding domain" can have an immunoglobulin heavy chain variable region comprising three heavy chain CDRs: CDR1, CDR2, and CDR3.

[0130] If the antibody or binding domain is expressed in 10 or more 5 M -1 Affinity or K a (i.e., the equilibrium association constant for a specific binding interaction, expressed in 1 / M) to a target, then it "specifically binds" to the target without significantly binding to other components present in the test sample. Antibodies or binding domains can be divided into "high affinity" antibodies or binding domains and "low affinity" antibodies or binding domains. "High affinity" antibodies or binding domains are those that have at least 10 7 M-1 , at least 10 8 M -1 , at least 10 9 M -1 , at least 10 10 M -1 , at least 10 11 M -1 , at least 10 12 M -1 or at least 10 13 M -1 K a "Low affinity" antibodies or binding domains are those that have a low affinity of up to 10 7 M -1 , up to 10 6 M -1 , up to 10 5 M -1 K a Alternatively, affinity can be defined as the equilibrium dissociation constant (K) for a specific binding interaction. d ), the unit is M (for example 10 -5 M to 10 -13 M). In the case of antibody binding to antigen, K a =1 / K d The affinity of an antibody or binding domain according to the present disclosure can be readily determined using conventional techniques such as surface plasmon resonance (see, e.g., Scatchard et al. (1949) Ann. NY Acad. Sci. 51:660; and U.S. Patent Nos. 5,283,173, 5,468,614, or equivalents).

[0131] As used herein, "CD152" refers to cluster of differentiation 152, which is also known as cytotoxic T lymphocyte-associated protein 4 (CTLA-4). The terms "CD152," "CTLA-4," and "CTLA4" are used interchangeably herein. Similarly, "anti-CD152," "anti-CTLA-4," and "anti-CTLA4" are also used interchangeably herein.

[0132] As used herein, "CD80" refers to cluster of differentiation 80, a protein found on dendritic cells, activated B cells, and monocytes that provides the co-stimulatory signal necessary for T cell activation and survival. The terms "CD80," "B7-1," and "B7.1" are used interchangeably herein.

[0133] As used herein, "CD86" refers to cluster of differentiation 86, a protein expressed on antigen-presenting cells that provides the co-stimulatory signals necessary for T cell activation and survival. The terms "CD86," "B7-2," and "B7.2" are used interchangeably herein.

[0134] As used herein, "conservative substitution" is considered in the art to be a substitution of one amino acid with another amino acid having similar properties. Exemplary conservative substitutions are well known in the art (see, for example, WO 97 / 09433, page 10, published on March 13, 1997; Lehninger, Biochemistry, Second Edition; Worth Publishers, Inc. NY: NY (1975), pp. 71-77; Lewin, Genes IV, Oxford University Press, NY and Cell Press, Cambridge, MA (1990), p. 8). In certain embodiments, conservative substitutions include leucine to serine substitutions.

[0135] As used herein, "ipilimumab analog" refers to a monoclonal antibody that specifically binds to CTLA-4 and comprises a heavy chain having the amino acid sequence of SEQ ID NO: 199 and a light chain having the amino acid sequence of SEQ ID NO: 200.

[0136] As used herein, unless otherwise indicated, any nonproprietary or common name of a biological product includes the biological product and any biosimilar products thereof. For example, the nonproprietary name ipilimumab refers to the biological product sold under the trade name YERVOY; it also includes any biosimilar products of that biological product.

[0137] As used herein, and unless otherwise indicated, the term "biosimilar product" refers to a biological product that 1) has the same amino acid sequence as a reference product; 2) has an amino acid sequence that differs from that of the reference product (e.g., an N-terminal or C-terminal truncation); or 3) has a post-translational modification (e.g., glycosylation or phosphorylation) that differs from that of the reference product, wherein the biosimilar product and the reference product utilize the same one or more mechanisms of action to prevent, treat, or cure a disease or condition.

[0138] As used herein, the term "derivative" refers to the modification of one or more amino acid residues of a peptide by chemical or biological methods, with or without the use of enzymes, such as by glycosylation, alkylation, acylation, ester formation or amide formation.

[0139] As used herein, a polypeptide or amino acid sequence "derived from" a specified polypeptide or protein refers to the source of the polypeptide. In certain embodiments, a polypeptide or amino acid sequence derived from a particular sequence (sometimes referred to as a "starting" or "parent" or "parental" sequence) has an amino acid sequence that is substantially identical to the starting sequence or a portion thereof, wherein the portion consists of at least 10-20 amino acids, at least 20-30 amino acids, or at least 30-50 amino acids, or at least 50-150 amino acids, or is otherwise identifiable to one of ordinary skill in the art as originating from the starting sequence. For example, a binding domain can be derived from an antibody, e.g., Fab, F(ab')2, Fab', scFv, a single domain antibody (sdAb), or the like.

[0140] A polypeptide derived from another polypeptide may have one or more mutations relative to the starting polypeptide, for example, one or more amino acid residues have been substituted by another amino acid residue or it has one or more amino acid residue insertions or deletions. The polypeptide may comprise a non-naturally occurring amino acid sequence. Such variations will necessarily have less than 100% sequence identity or similarity to the starting polypeptide. In one embodiment, the variant will have an amino acid sequence identity or similarity of about 60% to less than 100% to the amino acid sequence of the starting polypeptide. In another embodiment, the variant will have an amino acid sequence identity or similarity of about 75% to less than 100%, about 80% to less than 100%, about 85% to less than 100%, about 90% to less than 100%, or about 95% to less than 100% to the amino acid sequence of the starting polypeptide.

[0141] As used herein, unless otherwise provided, the positions of amino acid residues in the variable region of an immunoglobulin molecule are numbered according to the IMGT numbering convention (Brochet, X, et al, Nucl. Acids Res. (2008) 36, W503-508) and the positions of amino acid residues in the constant region of an immunoglobulin molecule are numbered according to the EU nomenclature (Ward et al., 1995 Therap. Immunol. 2: 77-94). Other numbering conventions are known in the art (e.g., the Kabat numbering convention (Kabat, Sequences of Proteins of Immunological Interest, 5 th ed. Bethesda, MD: Public Health Service, National Institutes of Health (1991)).

[0142] As used herein, the term "human" antibody refers to an antibody of human origin or a humanized antibody.

[0143] As used herein, the term "humanization" refers to the process of reducing the immunogenicity of antibodies or immunoglobulin binding proteins and polypeptides derived from non-human species (e.g., mouse or rat) to humans using genetic engineering techniques while still retaining the antigen binding properties of the original antibody. In some embodiments, the binding domains (e.g., light and heavy chain variable regions, Fab, scFv) of antibodies or immunoglobulin binding proteins and polypeptides are humanized. A technique called CDR grafting (Jones et al., Nature 321:522 (1986)) and its variations, including "reshaping" (Verhoeyen, et al., 1988 Science 239:1534-1536; Riechmann, et al., 1988 Nature 332:323-337; Tempest, et al., Bio / Technol 1991 9:266-271), "hyperchimerization" (Queen, et al., 1989 Proc Natl Acad Sci USA 86:10029-10033; Co, et al., 1991 Proc Natl Acad Sci USA 88:2869-2873; Co, et al., 1992 J Immunol Humanization of non-human binding domains is accomplished by methods such as "humanization" (Mark, et al., "Derivation of therapeutically active humanized and veneered anti-CD18 antibodies." In: Metcalf BW, Dalton BJ, eds. Cellular adhesion: molecular definition to therapeutic potential. New York: Plenum Press, 1994: 291-312). Other regions of antibody or immunoglobulin binding proteins and polypeptides, such as hinge and constant region domains, can also be humanized if derived from non-human sources.

[0144] As used herein, the term "patient in need thereof" or "subject in need thereof" refers to a patient or subject at risk of, or suffering from, a disease, disorder or condition that is amenable to treatment or amelioration with a CD152-binding antibody or composition thereof provided herein.

[0145] As used herein, the term "pharmaceutically acceptable" refers to molecular entities and compositions that do not typically produce allergic or other serious adverse reactions when administered using routes well known in the art. Molecular entities and compositions that are approved by federal or state regulatory agencies or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans, are considered "pharmaceutically acceptable."

[0146] As used herein, the terms "treat" or "improve" refer to therapeutic treatment or prophylactic / preventative treatment. If at least one symptom of a disease is improved in the individual being treated, or if treatment can delay the worsening of a progressive disease in the individual, or prevent the onset of another related disease, then the treatment is therapeutic.

[0147] As used herein, the term "therapeutically effective amount (or dose)" or "effective amount (or dose)" of a specific binding molecule or compound refers to an amount of a compound sufficient to result in a statistically significant improvement in one or more symptoms of the disease being treated, or a statistically significant improvement in organ function. When referring to an individual active ingredient administered alone, the therapeutically effective dose refers to that ingredient alone. When referring to a combination, the therapeutically effective dose refers to the combined amounts of the active ingredients that result in a therapeutic effect, whether administered sequentially or simultaneously (in the same formulation or in separate formulations).

[0148] As used herein, the terms "antibody-dependent cell-mediated cytotoxicity" and "ADCC" refer to a cell-mediated process in which nonspecific cytotoxic cells expressing FcγRs (e.g., monocytes, such as natural killer (NK) cells and macrophages) recognize bound antibodies (or other proteins capable of binding to FcγRs) on target cells and subsequently cause target cell lysis. In principle, any effector cell with an activating FcγR can be triggered to mediate ADCC. The primary cells mediating ADCC are NK cells, which express only FcγRIII, while monocytes can express FcγRI, FcγRII, and FcγRIII depending on their activation, localization, or differentiation state. For a review of FcγR expression on hematopoietic cells, see, for example, Ravetch et al., 1991, Annu. Rev. Immunol., 9:457-92.

[0149] As used herein, the term "promoter" refers to a region of DNA involved in binding RNA polymerase to initiate transcription.

[0150] As used herein, the term "nucleic acid," "nucleic acid molecule," or "polynucleotide" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in single-stranded or double-stranded form. Unless otherwise limited, the term encompasses nucleic acids containing analogs of natural nucleotides that have similar binding properties to the reference nucleic acid and are metabolized in a manner similar to natural nucleotides. 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 can 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. (1991) Nucleic Acid Res. 19:5081; Ohtsuka et al. (1985) J. Biol. Chem. 260:2605-2608; Cassol et al. (1992); Rossolini et al. (1994) Mol. Cell. Probes 8:91-98). The term nucleic acid can be used interchangeably with gene, cDNA, and mRNA encoded by a gene. As used herein, the term "nucleic acid," "nucleic acid molecule," or "polynucleotide" is intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), analogs of DNA or RNA generated using nucleotide analogs, and derivatives, fragments, and homologs thereof.

[0151] The term "expression" refers to the biosynthesis of a product encoded by a nucleic acid. For example, where a nucleic acid segment encodes a polypeptide of interest, expression involves transcribing the nucleic acid segment into mRNA and translating the mRNA into one or more polypeptides.

[0152] The terms "expression unit" and "expression cassette" are used interchangeably herein and refer to a nucleic acid segment that encodes a polypeptide of interest and is capable of providing expression of the nucleic acid segment in a host cell. An expression unit typically comprises a transcriptional promoter, an open reading frame encoding the polypeptide of interest, and a transcriptional terminator, all in an operable configuration. In addition to the transcriptional promoter and terminator, an expression unit may further comprise other nucleic acid segments, such as, for example, an enhancer or a polyadenylation signal.

[0153] As used herein, the term "expression vector" refers to a linear or circular nucleic acid molecule comprising one or more expression units. In addition to one or more expression units, an expression vector may also include additional nucleic acid segments, such as, for example, one or more origins of replication or one or more selectable markers. Expression vectors are typically derived from plasmid or viral DNA, or may contain elements of both.

[0154] As used herein, the term "sequence identity" refers to the relationship between two or more polynucleotide sequences or two or more polypeptide sequences. When a position in a sequence is occupied by the same nucleic acid base or amino acid residue in the corresponding position of the comparison sequence, the sequence is called "identical" at that position. The percentage "sequence identity" is calculated by determining the number of positions at which the same nucleic acid base or amino acid residue appears in the two sequences to generate the number of "identical" positions. The number of "identical" positions is then divided by the total number of positions in the comparison window and multiplied by 100 to generate the percentage of "sequence identity". The percentage of "sequence identity" is determined by comparing the two best aligned sequences in the comparison window. The comparison window of nucleotide sequence can be for example at least 20,30,40,50,60,70,80,90,100,110,120,130,140,150,160,170,180,190,200,300,400,500,600,700,800,900 or 1000 or more nucleic acids in length.The comparison window of peptide sequence can be for example at least 20,30,40,50,60,70,80,90,100,110,120,130,140,150,160,170,180,190,200,300 or more amino acid in length.In order to optimally align sequence to compare, the part of polynucleotide or peptide sequence in the comparison window can comprise the addition or disappearance that is called room, and reference sequence keeps constant. An optimal alignment is one that produces the maximum possible number of "identical" positions between the reference and comparison sequences, even with gaps. The percentage "sequence identity" between two sequences can be determined using a version of the program "BLAST 2 Sequences," available from the National Center for Biotechnology Information as of September 1, 2004, which includes the programs BLASTN (for nucleotide sequence comparisons) and BLASTP (for polypeptide sequence comparisons), which are based on the algorithm of Karlin and Altschul (Proc. Natl. Acad. Sci. USA 90(12):5873-5877, 1993). When using "BLAST 2 Sequences," the parameters that were used as default parameters as of September 1, 2004, can be used for word length (3), open gap penalty (11), extension gap penalty (1), gap reduction (50), expectation (10), and any other necessary parameters, including but not limited to the matrix option.Two nucleotide or amino acid sequences are said to have "substantially similar sequence identity" or "substantial sequence identity" if the two sequences have at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity relative to each other.

[0155] Antibody

[0156] Disclosed herein are human monoclonal antibodies comprising a CD152 binding domain. The antibody can be a heavy chain-only antibody. The antibody can be composed of only two heavy chains. The antibody can contain no light chains. The antibody can specifically bind to CD152. The antibody can be an isolated monoclonal antibody that specifically binds to CD152 with high affinity.

[0157] The anti-CD152 antibodies disclosed herein can specifically bind to human CD152. In some cases, the anti-CD152 antibodies can bind to human CD152 (e.g., K D <6.0*10 -11 M). When compared to ipilimumab analogs, anti-CD152 antibodies can have an affinity comparable to or higher than CTLA-4. Anti-CD152 antibodies can also block the binding of CD152 to its ligand B7.1. Anti-CD152 antibodies can have an enhanced tumor / peripheral serum ratio than ipilimumab analogs. Anti-CD152 antibodies can induce higher ADCC, for example, the antibody can induce an increase in NK cell lytic activity by at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 15 times, or at least about 20 times.

[0158] Anti-CD152 antibodies can include a CD152 binding domain comprising an immunoglobulin heavy chain variable region comprising CDR1, CDR2, and CDR3. Anti-CD152 antibodies can also include CDR1, CDR2, and CDR3 that differ from the CDR1, CDR2, and CDR3 of anti-CD152 antibodies disclosed herein by one or more conservative modifications. It will be understood in the art that certain conservative sequence modifications can be made without removing antigen binding. See, e.g., Brummell et al., 1993, Biochem 32:1180-8; de Wildt et al., 1997, Prot. Eng. 10:835-41; Komissrov et al., 1997, J. Biol. Chem. 272:26864-26870; Hall et al., 1992, J. Immunol. 149:1605-12; Kelley and O'Connell, 1993, Biochem. 32:6862-35; Adib-Conquy et al., 1998, Int. Immunol. 10:341-6 and Beers et al., 2000, Clin. Can. Res. 6:2835-43.

[0159] Pharmaceutical compositions and formulations

[0160] The pharmaceutical composition may comprise one or more anti-CD152 antibodies disclosed herein formulated with a pharmaceutically acceptable excipient. If the administration of an excipient can be tolerated by the recipient patient, it is referred to as a "pharmaceutically acceptable excipient". Excipients that can be used include carriers, surfactants, thickeners or emulsifiers, solid binders, dispersion or suspension aids, solubilizers, colorants, flavorings, coatings, disintegrants, lubricants, sweeteners, preservatives, isotonic agents, and combinations thereof. The selection and use of suitable excipients are taught in Gennaro, ed., Remington: The Science and Practice of Pharmacy, 20th Ed. (Lippincott Williams & Wilkins 2003), and Gennaro, ed., Remington's Pharmaceutical Sciences (Mack Publishing Company, 19th ed. 1995). Sterile phosphate-buffered saline is an example of a pharmaceutically acceptable excipient. The formulation may further include one or more carriers, diluents, preservatives, solubilizers, buffers, albumin to prevent protein loss from vial surfaces, and the like.

[0161] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form can vary depending on the subject being treated and the particular mode of administration, and generally can be that amount of the pharmaceutical composition that produces a therapeutic effect. Generally, the amount of active ingredient can range from about 0.01% to about 99% (w / w) of the composition, for example, can be about 0.1%-1%, about 0.1%-5%, about 0.1-10%, about 0.1%-20%, about 0.5%-1%, about 0.5%-5%, about 0.5%-10%, about 0.5%-20%, about 1%-5%, about 1%-10%, about 1%-20%, about 5%-10%, about 5%-20%, about Preferably, the amount of active ingredient can be from about 0.1% to about 70%, and most preferably from about 1% to about 30% of the pharmaceutical composition.

[0162] The pharmaceutical composition can be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active ingredient can be coated in a substance to protect it from the effects of acid and other natural conditions that may inactivate it. As used herein, the phrase "parenteral administration" means a mode of administration other than enteral and topical administration, typically by injection, and includes but is not limited to intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardial, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. Alternatively, the antibodies of the present invention can be administered via non-parenteral routes, such as topical, epidermal or mucosal administration routes, for example, intranasal, oral, vaginal, rectal, sublingual or topical. The pharmaceutical composition can be in the form of a sterile aqueous solution or dispersion. The pharmaceutical composition can also be formulated into microemulsions, liposomes or other ordered structures suitable for high drug concentrations.

[0163] The pharmaceutical composition can be formulated into a dosage form selected from the group consisting of oral unit dosage forms, intravenous unit dosage forms, intranasal unit dosage forms, suppository unit dosage forms, intradermal unit dosage forms, intramuscular unit dosage forms, intraperitoneal unit dosage forms, subcutaneous unit dosage forms, epidural unit dosage forms, sublingual unit dosage forms, and intracerebral unit dosage forms. The oral unit dosage form can be selected from the group consisting of tablets, pills, pellets, capsules, powders, lozenges, granules, solutions, suspensions, emulsions, syrups, elixirs, sustained-release formulations, aerosols, and sprays.

[0164] The pharmaceutical composition can be a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0165] The monoclonal antibodies disclosed herein can be formulated to ensure proper distribution in vivo. For example, to ensure that the therapeutic antibodies of the invention cross the blood-brain barrier, they can be formulated into liposomes, which can additionally contain targeting moieties to enhance selective transport to specific cells or organs. See, e.g., U.S. Patent Nos. 4,522,811; 5,374,548; 5,416,016; and 5,399,331; VV Ranade, 1989, J. Clin. Pharmacol. 29:685; Umezawa et al., 1988, Biochem. Biophys. Res. Commun. 153:1038; Bloeman et al., 1995, FEBS Lett. 357:140; M. Owais et al., 1995, Antimicrob. Agents Chemother. 39:180; Briscoe et al., 1995, Am. J. Physiol. 1233:134; Schreier et al., 1994, J. Biol. Chem. 269:9090; Keinanen and Laukkanen, 1994, FEBS Lett. 346:123; and Killion and Fidler, 1994, Immunomethods 4:273.

[0166] The pharmaceutical composition may optionally contain one or more additional pharmaceutically active ingredients, such as another antibody or a drug.The pharmaceutical composition of the invention may also be administered in combination therapy with, for example, another anticancer agent, another anti-inflammatory agent or a vaccine.

[0167] The pharmaceutical composition can be provided as a kit comprising a container comprising the pharmaceutical composition as described herein. The pharmaceutical composition can be provided, for example, in the form of an injectable solution for single or multiple doses, or as a sterile powder to be reconstituted prior to injection. Alternatively, such a kit can include a dry powder disperser, a liquid aerosol generator, or a nebulizer for administering the pharmaceutical composition. Such a kit can further include written information about the indications and usage of the pharmaceutical composition.

[0168] Treatment

[0169] Further disclosed herein is a method for treating a disease by administering a therapeutically effective amount of an antibody or pharmaceutical composition disclosed herein to a subject. The anti-CD152 antibodies disclosed herein can be used in a method for treating a subject (e.g., a human or non-human primate) or in a method for preparing a medicine for treating a subject. Typically, such methods include administering an anti-CD152 antibody as described herein to a subject in need of such treatment.

[0170] Anti-CD152 antibodies disclosed herein can be used in methods for treating subjects (e.g., humans or non-human primates) or for preparing methods for treating subjects. Typically, such methods include administering an anti-CD152 antibody as described herein to a subject in need of such treatment. In some embodiments, the anti-CD152 antibody comprises at least one effector function selected from antibody-dependent cell-mediated cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), such that the anti-CD152 antibody induces ADCC and / or CDC for cells expressing CD152 in the subject.

[0171] Also disclosed herein is a method for treating a disease (such as cancer) characterized by overexpression of a tumor antigen. Examples of tumor antigens that can be identified by bispecific anti-CD152 antibodies can include PSMA, CD19, CD20, CD37, CD38, CD123, Her2, ROR1, RON, glycoprotein A33 antigen (gpA33) and CEA. Typically, such methods include administering a therapeutically effective amount of an anti-CD152 antibody to a subject in need of such treatment, the CD152 antibody comprising a second binding domain in combination with a tumor antigen as described herein. Anti-CD152 antibodies can induce T cell cytotoxicity (RTCC) directed against cells expressing tumor antigens in a subject.

[0172] The methods can be used to treat cancers such as prostate cancer, colorectal cancer, renal cell carcinoma, bladder cancer, salivary gland cancer, pancreatic cancer, ovarian cancer, non-small cell lung cancer, melanoma, breast cancer (e.g., triple-negative breast cancer), adrenal cancer, mantle cell lymphoma, acute lymphoblastic leukemia, chronic lymphocytic leukemia, non-Hodgkin lymphoma, acute myeloid leukemia (AML), B-lymphoid leukemia, blastic plasmacytoid dendritic neoplasm (BPDCN), and hairy cell leukemia.

[0173] Also disclosed herein are methods of treating an autoimmune disorder comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition or an anti-CD152 antibody described herein.

[0174] The subject for administering an anti-CD152 antibody as described herein includes patients at high risk of developing a particular condition and patients showing the presence of such a condition. Typically, the subject has been diagnosed as having the condition for which treatment is sought. In addition, any changes in the subject's condition (e.g., an increase or decrease in the clinical symptoms of the condition) can be monitored during the course of treatment. Similarly, in some variations, the subject does not suffer from another condition that requires treatment involving cells that target CD152.

[0175] In prophylactic applications, the pharmaceutical composition can be administered to a patient susceptible to or at risk of a particular condition in an amount sufficient to eliminate or reduce the risk of the condition or to delay the onset of the condition. In therapeutic applications, the composition can be administered to a patient suspected of or already suffering from such a condition in an amount sufficient to cure or at least partially suppress the symptoms of the condition and its complications. An amount sufficient to accomplish this is referred to as a therapeutically effective dose or therapeutically effective amount. In both prophylactic and therapeutic regimens, the medicament can be administered in several doses until a sufficient response is achieved. Typically, if the desired response begins to weaken, the response is monitored and repeated doses are given.

[0176] In order to identify a subject patient for treatment according to the methods of the present disclosure, recognized screening methods can be used to determine risk factors associated with a specific condition or to determine the status of an identified pre-existing condition in the subject. Such methods can include, for example, determining whether an individual has a relative diagnosed with a specific condition. Screening methods can also include, for example, routine examinations to determine the familial status of a specific condition known to have a genetic component. For example, it is also known that various cancers have certain heritable components. Heritable components of cancer include, for example, mutations in multiple genes that are being transformed (e.g., Ras, Raf, EGFR, cMet, etc.), the presence or absence of certain HLA and killer inhibitory receptor (KIR) molecules, or the mechanism by which cancer cells can directly or indirectly regulate the immunosuppression of cells such as NK cells and T cells (see, for example, Ljunggren and Malmberg, Nature Rev. Immunol. 7:329-339, 2007; Boyton and Altmann, Clin. Exp. Immunol. 149:1-8, 2007). For this purpose, nucleotide probes can be routinely used to identify individuals carrying genetic markers associated with a specific condition of interest. In addition, a variety of immunological methods are known in the art that can be used to identify markers for specific conditions. For example, various ELISA immunoassays employing monoclonal antibody probes to detect antigens associated with specific tumors are available and well known in the art. Screening can be performed as indicated by known patient symptoms, age factors, associated risk factors, and the like. These methods allow clinicians to routinely select patients who require treatment with the methods described herein. According to these methods, targeting cells that pathologically express tumor antigens can be implemented as an independent treatment program or as a follow-up, adjuvant, or coordinated treatment regimen for other treatments.

[0177] In this case, the determination of the effective dose is generally based on animal model studies, followed by human clinical trials, and is guided by determining an effective dose and administration regimen that significantly reduces the occurrence or severity of the subject's condition in the model subject. The effective dose of the composition of the present disclosure varies depending on many different factors, including the mode of administration, the target site, the patient's physiological state, whether the patient is human or animal, other drugs administered, whether the treatment is prophylactic or therapeutic, and the specific activity of the composition itself and its ability to elicit the desired response in the individual. Typically, the patient is a human, but in some diseases, the patient can be a non-human mammal. Typically, the dosage regimen is adjusted to provide the best therapeutic response, i.e., to optimize safety and efficacy. Therefore, a therapeutically effective amount is also an amount in which the beneficial effects of administering an anti-CD152 antibody as described herein outweigh any undesirable side effects. For administration of an anti-CD152 antibody, the dosage range can be from about 0.1 μg to 100 mg / kg or 1 μg / kg to about 50 mg / kg of the subject's body weight, and more typically 10 μg to 5 mg / kg. In more specific embodiments, the effective amount of the medicament is between about 1 μg / kg and about 20 mg / kg, between about 10 μg / kg and about 10 mg / kg or between about 0.1 mg / kg and about 5 mg / kg. Dosages within this range can be achieved by single or multiple administrations, including, for example, multiple administrations per day or daily, weekly, biweekly or monthly administrations. For example, in some variations, the regimen consists of an initial administration, followed by multiple subsequent administrations at weekly or biweekly intervals. Another regimen consists of an initial administration, followed by multiple subsequent administrations at monthly or bimonthly intervals. Alternatively, it can be administered irregularly as indicated by monitoring the clinical symptoms of the disease.

[0178] The doctor in charge can change the dosage of pharmaceutical composition, to maintain the concentration of expectation at target site.For example, if the mode of intravenous delivery is selected, depending on the state of experimenter and the measurement reaction of expectation, the local concentration of medicament in target tissue place blood flow can be between about 0.01-50 nanomoles of every liter of composition, sometimes between about 1.0 nanomoles and 10,15 or 25 nanomoles of every liter of every liter.Can select higher or lower concentration based on the mode of delivery (for example, percutaneous delivery and delivery to mucosal surface).Dosage should also be adjusted based on the release rate of the formulation (for example, nasal spray and powder, slow-release oral or injection granules, transdermal formulations etc.) used.To reach identical serum concentration level, for example, the slow-release granules (under standard conditions) with 5 nanomoles of release rate will be used with the granules with 10 nanomoles of release rate of about twice dosage.

[0179] Anti-CD152 therapeutic agents (e.g., anti-CD152 antibodies) can also be administered at a daily dose of approximately 0.001 to approximately 10 milligrams (mg) per kilogram of body weight (mpk), preferably in a single daily dose or in divided doses of approximately twice to six times per day. In order to administer to human adult patients, a therapeutically effective amount can be administered with a dosage in the range of 0.2mg to 800mg per dose, including but not limited to 0.2mg per dose, 0.5mg per dose, 1mg per dose, 5mg per dose, 10mg per dose, 25mg per dose, 100mg per dose, 200mg per dose, and 400mg per dose, and can be administered repeatedly during treatment, typically continuous daily doses. Anti-CD152 therapeutic agents can be administered at different times of the day. In one embodiment, the optimal therapeutic dose can be administered at night. In another embodiment, the optimal therapeutic dose can be administered in the morning. Thus, in one embodiment, the total daily dose of the anti-CD152 therapeutic may range from about 1 mg to about 2 g, and typically ranges from about 100 mg to about 1.5 g, and most typically ranges from about 200 mg to about 1200 mg. In the case of a typical 70 kg adult human, the total daily dose of the anti-CD152 therapeutic may range from about 2 mg to about 1200 mg, and as described above will typically range from about 0.2 mg to about 800 mg.

[0180] The dosage regimen may also be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the urgency of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniform dosage. As used herein, dosage unit form refers to physically discrete units suitable as unit doses for a subject to be treated; each unit contains a predetermined amount of active ingredient calculated to produce the desired therapeutic effect together with the required pharmaceutical carrier. Alternatively, the antibody may be administered as a sustained-release formulation, in which case less frequent administration is required.

[0181] For administration of antibodies, the range of dosage can be about 0.0001 to 100 mg / kg of the host's body weight, and more generally 0.01 to 5 mg / kg. For example, the dosage can be 0.3 mg / kg body weight, 1 mg / kg body weight, 3 mg / kg body weight, 5 mg / kg body weight or 10 mg / kg body weight or in the range of 1-10 mg / kg. Exemplary treatment regimens require administration once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every three months or once every three to six months. Preferred dosage regimens for the anti-CD152 antibodies of the present invention include 1 mg / kg body weight or 3 mg / kg body weight administered intravenously, wherein the antibody is administered using one of the following dosing regimens: (i) six doses every four weeks, then every three months; (ii) every three weeks; (iii) 3 mg / kg body weight once, then 1 mg / kg body weight every three weeks. In some methods, the dosage is adjusted to achieve a plasma antibody concentration of about 1-1000 μg / ml, and in some methods, about 25-300 μg / ml.

[0182] A "therapeutically effective dose" of an anti-CD152 antibody of the present invention preferably results in a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or prevention of impairment or disability due to disease distress. For example, for treating a tumor-bearing subject, a "therapeutically effective dose" preferably inhibits tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and still more preferably at least about 80%, relative to an untreated subject. A therapeutically effective amount of a therapeutic antibody can reduce tumor size or otherwise improve symptoms in a subject, which is typically a human or can be another mammal.

[0183] Regarding the treatment of solid tumors, the scheme for evaluating endpoints and anti-tumor activity is well known in the art. Although each scheme may have a different definition for tumor response assessment, the RECIST (Effective Evaluation Criteria for Solid Tumors) standard is currently considered by the National Cancer Institute (National Cancer Institute) as a recommended guideline for evaluating tumor response (see Therasse et al., J. Natl. Cancer Inst. 92: 205-216, 2000). According to the RECIST standard, tumor response means reducing or eliminating all measurable lesions or metastases. If the disease includes a lesion that can be accurately measured as ≥20mm in at least one dimension with conventional technology, or a lesion that can be accurately measured as ≥10mm in at least one dimension with a spiral CT scan with a clearly defined edge by medical photos or X-rays, computer axial tomography (CT), magnetic resonance imaging (MRI) or clinical examination (if the lesion is superficial), then it is generally considered that the disease is measurable. Unmeasurable disease means that the disease includes lesions that are <20mm with conventional technology or <10mm with spiral CT scans and truly unmeasurable lesions (too small to be accurately measured). Unmeasurable disease included pleural effusion, ascites, and disease documented by indirect evidence.

[0184] The protocol requires objective status criteria to assess solid tumor response. Representative criteria include the following: (1) complete response (CR), defined as complete disappearance of all measurable disease; no new lesions; no disease-related symptoms; no evidence of measurable disease; (2) partial response (PR), defined as a 30% decrease in the sum of the longest diameters of target lesions; (3) progressive disease (PD), defined as a 20% increase in the sum of the longest diameters of target lesions or the appearance of any new lesions; (4) stable or non-response, defined as not meeting the criteria for CR, PR, or progressive disease. (See Therasse et al., supra)

[0185] Additional endpoints accepted in the oncology field include overall survival (OS), disease-free survival (DFS), objective response rate (ORR), time to progression (TTP), and progression-free survival (PFS) (see Guidance for Industry: Clinical Trial Endpoints for the Approval of Cancer Drugs and Biologics, April 2005, Center for Drug Evaluation and Research, FDA, Rockville, MD).

[0186] As monotherapy or in combination with anti-PD-L1 monoclonal antibodies or other anticancer drugs, anti-CD152 antibodies can be used to inhibit CTLA-4-mediated signaling pathways that negatively regulate immune responses and thereby enhance tumor-specific immune responses.

[0187] Methods for preparing antibodies

[0188] The antibodies disclosed herein can be human heavy chain-only antibodies (HCAbs) generated from Harbour humanized mice (U.S. Patent Nos. 9,353,179, 9,346,877, and 8,921,522, and European Patent Nos. 1776383 and 1864998). The molecules produced by HCAb mice can be soluble and can have affinity, diversity, and / or physicochemical properties comparable to traditional human IgG antibodies.

[0189] Preparation of HCAbs from HCAb mice can facilitate the generation of soluble human VH domains, the smallest immunoglobulin recognition unit, and thus the construction of new multifunctional molecules containing multiple VH domains or VH domains coupled to other molecules (such as bispecific antibodies, antibody-drug conjugates, or VH domain-derived diagnostic or therapeutic molecules).

[0190] One or more V-linked antibodies with the anti-CD152 antibodies disclosed herein may also be used. H Anti-CD152 antibodies can be prepared by modifying the variable region (i.e., V H and / or V L ) within one or more CDR regions and / or within one or more framework regions to engineer the antibody. Additionally or alternatively, an antibody can be engineered by modifying residues within the constant region, for example, to alter the effector functions of the antibody.

[0191] Polynucleotide molecules comprising the desired polynucleotide sequence can be propagated by placing the molecule in a vector. Viral and non-viral vectors, including plasmids, can be used. The choice of plasmid will depend on the type of cells in which propagation is desired and the purpose of the propagation. Certain vectors can be used to amplify and prepare large quantities of the desired DNA sequence. Other vectors are suitable for expression in cultured cells. Other vectors are also suitable for transfer and expression in cells in whole animals or humans. The selection of an appropriate vector is entirely within the skill of the art. Many such vectors are commercially available. Partial or full-length polynucleotides are typically inserted into a vector by attaching DNA ligase to restriction enzyme sites cut into the vector. Alternatively, the desired nucleotide sequence can be inserted by homologous recombination in vivo. Typically, this is achieved by attaching regions of homology to the vector on the sides of the desired nucleotide sequence. For example, regions of homology are added by ligation of oligonucleotides or by polymerase chain reaction using primers containing both regions of homology and a portion of the desired nucleotide sequence.

[0192] For expression, an expression cassette or system can be used. In order to express the nucleic acid encoding the polypeptide disclosed herein, a nucleic acid molecule encoding the polypeptide operably connected to a regulatory sequence that controls transcriptional expression in an expression vector is introduced into a host cell. In addition to transcriptional regulatory sequences such as promoters and enhancers, the expression vector can also include translational regulatory sequences and marker genes (which are suitable for selecting cells carrying the expression vector). The gene products encoded by the polynucleotides of the present disclosure are expressed in any convenient expression system, including, for example, bacterial, yeast, insect, amphibian, and mammalian systems. In the expression vector, the polynucleotide encoding the polypeptide is appropriately connected to the regulatory sequence to obtain the desired expression characteristics. These can include promoters, enhancers, terminators, operators, repressors, and inducers. The promoter can be regulated (e.g., a promoter from a steroid-inducible pIND vector (Invitrogen)) or constitutive (e.g., a promoter from CMV, SV40, an elongation factor, or an LTR sequence). They are connected to the desired nucleotide sequence using the techniques described above for connection to the vector. Any technique known in the art can be used. Thus, expression vectors will generally provide a transcriptional and translational initiation region, which may be inducible or constitutive, with the coding region operably linked under the transcriptional control of the transcriptional initiation region and a transcriptional and translational termination region.

[0193] The expression cassette can be introduced into a variety of vectors, such as plasmids, BACs, YACs, phages such as lambda, P1, M13, plant or animal virus vectors (such as retroviral vectors, adenovirus vectors), etc., wherein the feature of the vector is generally to provide the ability to select cells comprising the expression vector. The vector can provide extrachromosomal maintenance, particularly as a plasmid or virus, or provide integration into the host chromosome. In the case of the desired extrachromosomal maintenance, a starting point sequence for plasmid replication is provided, which can be a low copy number or a high copy number. A variety of markers can be used for selection, particularly markers for defense toxins, more particularly markers for defense antibiotics. The specific markers selected are selected according to the properties of the host, wherein in some cases, can be complemented with auxotrophic hosts. The guidance of the DNA construct can use any convenient method, including, for example, conjugation, bacterial transformation, calcium-precipitated DNA, electroporation, fusion, transfection, infection with viral vectors, gene guns (biolistics), etc. The present disclosure relates to expression vectors comprising a nucleic acid segment, wherein the nucleic acid segment may comprise the nucleotides set forth in SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 163, 169, 175, 181, or 187.

[0194] Thus, the proteins used in the present disclosure can be produced in genetically engineered host cells according to conventional techniques. Suitable host cells are those cell types that can be transformed or transfected with exogenous DNA and grown in culture, and include bacteria, fungal cells, and cultured higher eukaryotic cells (including cultured cells of multicellular organisms), particularly cultured mammalian cells. Techniques for manipulating cloned DNA molecules and introducing exogenous DNA into various host cells are disclosed in Sambrook and Russell, Molecular Cloning: A Laboratory Manual (3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2001) and Ausubel et al., Short Protocols in Molecular Biology (4th ed., John Wiley & Sons, 1999).

[0195] In order to guide the recombinant protein into the secretory pathway of the host cell, a secretory signal sequence (also referred to as a leader sequence) is provided in the expression vector. The secretory signal sequence can be a native form of the recombinant protein, or can be derived from another secretory protein or synthesized de novo. The secretory signal sequence is operably connected to the DNA sequence encoding the polypeptide, that is, the two sequences are joined and positioned in the correct reading frame to guide the newly synthesized polypeptide into the secretory pathway of the host cell. The secretory signal sequence is generally located 5' of the DNA sequence encoding the polypeptide of interest, although some signal sequences can be located at other positions of the DNA sequence of interest (see, for example, Welch et al., U.S. Patent number 5,037,743; Holland et al., U.S. Patent number 5,143,830).

[0196] Cultured mammalian cells are suitable hosts for producing recombinant proteins for use in the present disclosure. Methods for introducing exogenous DNA into mammalian host cells include calcium phosphate-mediated transfection (Wigler et al., Cell 14:725, 1978; Corsaro and Pearson, Somatic Cell Genetics 7:603, 1981; Graham and Van der Eb, Virology 52:456, 1973), electroporation (Neumann et al., EMBO J. 1:841-845, 1982), DEAE-dextran-mediated transfection (Ausubel et al., supra), and liposome-mediated transfection (Hawley-Nelson et al., Focus 15:73, 1993; Ciccarone et al., Focus 15:80, 1993). Production of recombinant polypeptides in cultured mammalian cells is disclosed in, for example, Levinson et al., US Patent No. 4,713,339; Hagen et al., US Patent No. 4,784,950; Palmiter et al., US Patent No. 4,579,821; and Ringold, US Patent No. 4,656,134. Examples of suitable mammalian host cells include African green monkey kidney cells (Vero; ATCC CRL 1587), human embryonic kidney cells (293-HEK; ATCC CRL 1573), baby hamster kidney cells (BHK-21, BHK-570; ATCC CRL 8544, ATCC CRL 10314), canine kidney cells (MDCK; ATCC CCL 34), Chinese hamster ovary cells (CHO-K1; ATCC CCL61; CHODG44; CHO DXB11 (Hyclone, Logan, UT); see also, e.g., Chasin et al., Som. Cell. Molec. Genet. 12:555, 1986), rat pituitary cells (GH1; ATCC CCL82), HeLa S3 cells (ATCC CCL2.2), rat hepatoma cells (H-4-II-E; ATCC CRL 34). 1548), SV40-transformed monkey kidney cells (COS-1; ATCC CRL 1650), and mouse embryonic cells (NIH-3T3; ATCC CRL 1658). Other suitable cell lines are known in the art and are available from public depositories, such as the American Type Culture Collection, Manassas, Virginia. Strong transcriptional promoters, such as those from SV-40 or cytomegalovirus, can be used. See, for example, U.S. Patent No. 4,956,288.Other suitable promoters include the promoter from the metallothionein gene (US Pat. Nos. 4,579,821 and 4,601,978) and the adenovirus major late promoter.

[0197] Drug selection is commonly used to select cultured mammalian cells that have been inserted with foreign DNA. Such cells are commonly referred to as "transfectants". Cells that are cultured in the presence of a selection agent and that are capable of delivering a gene of interest to their progeny are referred to as "stable transfectants". Exemplary selection markers include genes encoding resistance to the antibiotic neomycin, which allows selection in the presence of neomycin-type drugs such as G-418, etc.; the gpt gene of xanthine guanine phosphoribosyltransferase, which allows host cell growth in the presence of mycophenolic acid / xanthine; and markers providing resistance to zeocin, bleomycin, blastocidin, and hygromycin (see, for example, Gatignol et al., Mol. Gen. Genet. 207: 342, 1987; Drocourt et al., Nucl. Acids Res. 18: 4009, 1990). Selection systems can also be used to increase the expression level of a gene of interest, a process known as "amplification". Amplification is performed by culturing the transfectants in the presence of a low level of a selection agent and then increasing the amount of the selection agent to select for cells that produce high levels of the introduced gene product. An exemplary amplifiable selectable marker is dihydrofolate reductase, which confers resistance to methotrexate. Other drug resistance genes (e.g., hygromycin resistance, multidrug resistance, puromycin acetyltransferase) can also be used.

[0198] Other higher eukaryotic cells can also be used as hosts, including insect cells, plant cells, and avian cells. The use of Agrobacterium rhizogenes as a vector for expressing genes in plant cells has been reviewed by Sinkar et al., J. Biosci. (Bangalore) 11:47-58, 1987. Transformation of insect cells and the production of foreign polypeptides therein are disclosed by Guarino et al., US 5,162,222 and WO 94 / 06463.

[0199] Insect cells can be infected with recombinant baculovirus, which is generally derived from Autographa californica nuclear polyhedrosis virus (AcNPV). See King and Possee, The Baculovirus Expression System: A Laboratory Guide (Chapman & Hall, London); O'Reilly et al., Baculovirus Expression Vectors: A Laboratory Manual (Oxford University Press., New York 1994); and Baculovirus Expression Protocols. Methods in Molecular Biology (Richardson ed., Humana Press, Totowa, NJ, 1995). Recombinant baculovirus can also be produced by using a transposon-based system described by Luckow et al. (J. Virol. 67: 4566-4579, 1993). This system utilizing a transfer vector is commercially available in the form of a kit (BAC-TO-BAC kit; Life Technologies, Gaithersburg, MD). Transfer vectors (e.g., PFASTBAC1; Life Technologies) contain the Tn7 transposon to transfer the DNA encoding the protein of interest into the baculovirus genome maintained in E. coli as a large plasmid called a "bacmid." See Hill-Perkins and Possee, J. Gen. Virol. 71:971-976, 1990; Bonning et al., J. Gen. Virol. 75:1551-1556, 1994; and Chazenbalk and Rapoport, J. Biol. Chem. 270:1543-1549, 1995. In addition, the transfer vector can include an in-frame fusion with DNA encoding a polypeptide extension or affinity tag as described above. Using techniques known in the art, the transfer vector containing the protein-encoding DNA sequence is transformed into E. coli host cells, and the cells are screened for bacmids containing the interrupted lacZ gene, indicating the recombinant baculovirus. Bacmid DNA containing the recombinant baculovirus genome is isolated using conventional techniques and used to transfect Spodoptera frugiperda cells, such as Sf9 cells. Recombinant virus expressing the protein of interest is subsequently produced. Recombinant virus stocks are prepared using methods commonly used in the art.

[0200] To produce the protein, the recombinant virus is used to infect host cells, typically cell lines derived from fall armyworm, Spodoptera frugiperda (e.g., Sf9 or Sf21 cells) or Trichoplusia ni (e.g., HIGH FIVE cells; Invitrogen, Carlsbad, CA). See generally Glick and Pasternak, Molecular Biotechnology, Principles & Applications of Recombinant DNA (ASM Press, Washington, DC, 1994). See also U.S. Patent No. 5,300,435. Serum-free medium is used to grow and maintain the cells. Suitable medium formulations are known in the art and can be obtained from commercial suppliers. The cells are grown from approximately 2-5 x 10 5 Grow cells to a seeding density of 1-2 x 10 6 The cells are cultured at a density of 10 cells / mL, at which point the recombinant virus stock is added at a multiplicity of infection (MOI) of 0.1 to 10 (more typically close to 3). The protocols used are generally described in available laboratory manuals (see, e.g., King and Possee, supra; O'Reilly et al., supra; Richardson, supra).

[0201] Fungal cells, including yeast cells, can also be used in the present disclosure. In this regard, yeast species include, for example, Saccharomyces cerevisiae, Pichia pastoris, and Pichia methanolica. Methods for transforming Saccharomyces cerevisiae cells with exogenous DNA and producing recombinant polypeptides therefrom are disclosed in, for example, Kawasaki, U.S. Patent No. 4,599,311; Kawasaki et al., U.S. Patent No. 4,931,373; Brake, U.S. Patent No. 4,870,008; Welch et al., U.S. Patent No. 5,037,743; and Murray et al., U.S. Patent No. 4,845,075. Transformed cells are selected by a phenotype determined by a selectable marker, which is typically drug resistance or the ability to grow in the absence of a specific nutrient (e.g., leucine). The exemplary vector system used in Saccharomyces cerevisiae is the POT1 vector system disclosed by Kawasaki et al. (U.S. Patent No. 4,931,373), which allows the selection of transformed cells by growth in a medium containing glucose. Suitable promoters and terminators for use in yeast include promoters and terminators from glycolytic enzyme genes (see, for example, Kawasaki, U.S. Patent No. 4,599,311; Kingsman et al., U.S. Patent No. 4,615,974; and Bitter, U.S. Patent No. 4,977,092) and promoters and terminators from alcohol dehydrogenase genes. See also U.S. Patent Nos. 4,990,446; 5,063,154; 5,139,936; and 4,661,454. Transformation systems for other yeasts are known in the art, including Hansenula polymorpha, Schizosaccharomyces pombe, Kluyveromyces lactis, Kluyveromyces fragilis, Ustilago maydis, Pichia pastoris, Pichia methanolica, Pichia guillermondii, and Candida maltosa. See, for example, Gleeson et al., J. Gen. Microbiol. 132:3459-3465, 1986; Cregg, U.S. Pat. No. 4,882,279; and Raymond et al., Yeast 14:11-23, 1998. Aspergillus cells can be used according to the method of McKnight et al., U.S. Pat. No. 4,935,349.Methods for transforming Acremonium chrysogenum are disclosed in Sumino et al., U.S. Patent No. 5,162,228. Methods for transforming Neurospora are disclosed in Lambowitz, U.S. Patent No. 4,486,533. Production of recombinant proteins in Pichia methanolica is disclosed in U.S. Patent Nos. 5,716,808; 5,736,383; 5,854,039; and 5,888,768.

[0202] Prokaryotic host cells, including bacterial strains of Escherichia coli, Bacillus and other genera, are also useful host cells in the present disclosure. The technology for transforming these hosts and expressing the foreign DNA sequence cloned therein is well known in the art (see, for example, Sambrook and Russell, supra). When expressing recombinant proteins in bacteria such as Escherichia coli, the protein can usually be retained in the cytoplasm as insoluble particles, or can be directed to the periplasmic space by bacterial secretion sequences. In the former case, the cells are lysed, and the particles are recovered and denatured using, for example, guanidine isothiocyanate or urea. The denatured protein can then be refolded and dimerized by diluting the denaturant, such as by dialyzing against a combination of urea solution and reduced and oxidized glutathione, and then by dialyzing against a buffered saline solution. Alternatively, the protein can be recovered in a soluble form from the cytoplasm and separated without using a denaturant. Proteins are recovered as aqueous extracts from cells in, for example, phosphate buffered saline. In order to capture the protein of interest, the extract is directly applied to a chromatographic medium, such as an immobilized antibody or a heparin-agarose column. Secreted proteins can be recovered from the periplasmic space in a soluble and functional form by disrupting the cells (e.g., by sonication or osmotic shock) to release the contents of the periplasmic space and recovering the protein, thereby eliminating the need for denaturation and refolding. Antibodies, including single-chain antibodies, can be produced in bacterial host cells according to known methods. See, for example, Bird et al., Science 242:423-426, 1988; Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883, 1988; and Pantoliano et al., Biochem. 30:10117-10125, 1991.

[0203] In some embodiments, the host cell of the present invention is transformed or transfected by culture medium containing the required other components of nutrients and selected host cell growth.A variety of suitable culture mediums include defined composition culture medium and composite culture medium, which are known in the art and generally include carbon source, nitrogen source, essential amino acid, vitamin and mineral substance.As required, culture medium can also contain components such as somatomedin or serum.Growth medium will usually be selected to contain the cell of the DNA of external source interpolation by the drug selection or the shortage of essential nutrients that for example are supplemented by the selectable marker carried on the expression vector or cotransfection in the host cell.

[0204] Anti-CD152 antibodies can be purified by conventional protein purification methods, typically by a combination of chromatographic techniques. See generally Affinity Chromatography: Principles & Methods (Pharmacia LKB Biotechnology, Uppsala, Sweden, 1988); Scopes, Protein Purification: Principles and Practice (Springer-Verlag, New York 1994). Proteins containing the immunoglobulin Fc region can be purified by affinity chromatography on immobilized protein A or protein G. Additional purification steps such as gel filtration can be used to obtain the desired level of purity or to provide desalting, buffer exchange, etc.

[0205] The following examples of the present invention are intended to further illustrate the nature of the present invention. It should be understood that the following examples do not limit the present invention, and the scope of the present invention is determined by the appended claims. Example

[0206] Example 1 - Generation of anti-CTLA-4 antibodies

[0207] Human CTLA-4-ECD protein (Acro Bio) was used as an immunogen to generate anti-CTLA-4 antibodies. The use of human immunoglobulin transgenic mouse technology to develop and produce human antibodies was first described by Abgenix (xeno mouse and Medarex (HuMab "mouse"); Lonberg et al., 1994, Nature, 368:856-859; Lonberg and Huszar, 1995, Internal Rev. Immunol., 13:65-93; Harding and Lonberg, 1995, Ann. NY Acad. Sci., 764:536-546).

[0208] HCAb mice were immunized three times with 20 mg / per mouse of human CTLA-4-ECD protein every two weeks, and six of them were immunized another five times with 44 mg / per mouse. Except for the first injection (where Stimune (Prionics) was used as an adjuvant), all booster immunizations were performed with Ribi adjuvant (Sigma Adjuvant System S6322-1VL). After immunization, single cell suspensions were isolated from mouse bone marrow, spleen, and lymph nodes. Mouse plasma cells were then isolated using a plasma cell isolation kit (Miltenyi, Cat. No. 130-092-530). In short, total RNA from mouse plasma cells was prepared and reverse transcribed into cDNA in a larger pool. The following primers were used to amplify human VH regions from cDNA.

[0209] Forward primer:

[0210] lib-3-23 / 53-S:5'-GTTGTCCAGTGTGAGGTGCAGCTG (SEQ ID NO: 193) and

[0211] lib-3-11-S:5'-GTGTCCAGTGTCAGGTGCAGCTG (SEQ ID NO:194)

[0212] Reverse primer:

[0213] mG1hrv:5'-GGCTTACAACCACAATCCCTGGGC(SEQ ID NO:195)

[0214] All amplified PCR fragments containing the VH domain were cloned into the mammalian expression vector pTT5. The resulting plasmids were transformed into bacteria (DH5α) by electroporation. The plasmids were replicated and purified, and then transfected into HEK 293 cells for antibody production.

[0215] 293 cells were incubated in 293 FreeStyle culture medium (12338018, Thermo) for 10 days, and the supernatant was screened by ELISA assay. Recombinant human CTLA4-his protein (Acro Bio) was diluted in PBS at a concentration of 2 μg / mL, and 100 μL of diluted CTLA-4-his protein per well was added to an ELISA microplate, which was incubated overnight at 4 ° C to coat the plate with recombinant protein. The plate was then blocked with ELISA blocking solution (containing 2% BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer, w / v) at 37 ° C for 2 hours and then incubated with the supernatant at 37 ° C for 1 hour. The plate was washed and incubated with horseradish peroxidase (HRP)-conjugated goat anti-human IgG (H+L) antibody (A18805, Life technologies) at 37 ° C for 1 hour. 100 μL of tetramethylbenzidine (TMB) was added and the plate was incubated at room temperature for 15 minutes. 50 μL of 1N HCl was added to stop the reaction. 35 positive clones showing clear staining were selected for further testing.

[0216] Thirty-five clones were sequenced, and nine clones with unique CDR3 sequences were selected from the 35 clones. The nucleic acid and amino acid sequences of these nine anti-CTLA-4 antibodies are summarized in Table 1. HCAb antibodies contain only two heavy chains.

[0217] Table 1. Nucleic acid and amino acid sequences of human anti-CTLA-4 antibodies

[0218]

[0219]

[0220] na: nucleic acid; aa: amino acid

[0221] Example 2 - Preparation and purification of anti-CTLA-4 antibodies

[0222] Step 1. Preparation of HEK 293F cells overexpressing hCTLA-4

[0223] The nucleotide sequence encoding human CTLA-4 (SEQ ID NO: 196, encoding the amino acid sequence of SEQ ID NO: 197) was subcloned into the pcDNA3.1 vector (Clontech) to obtain a plasmid. HEK293 and CHO-K1 cells (Invitrogen) were transiently transfected with the plasmid using PEI, and the transformants were cultured in DMEM medium containing 0.5 g / mL penicillin / streptomycin and 10% (w / w) fetal bovine serum (FBS) for 2 weeks. Limiting dilutions were performed in 96-well culture plates, and the plates were then incubated at 37° C. with 5% (v / v) CO 2 for approximately 2 weeks. Single clones were expanded in 6-well plates, and the expanded clones were screened by flow cytometry using a commercially available anti-hCTLA-4 antibody (R&D Systems). Clones that showed a higher growth rate and higher fluorescence intensity as measured by FACS were further expanded and cryopreserved in liquid nitrogen.

[0224] Step 2. Determine the binding activity of anti-CTLA-4 antibodies in HEK 293F cell culture medium by ELISA and cell-based FACS binding assays

[0225] Recombinant human CTLA4-his protein (Acro Bio) was diluted in PBS at a concentration of 2 μg / mL, and 100 μL of diluted CTLA-4-his protein per well was added to an ELISA microplate, which was incubated overnight at 4°C to coat the plate with the recombinant protein. The plate was then blocked with ELISA blocking solution (containing 2% BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer, w / v) at 37°C for 2 hours, and then incubated with 293F cell culture medium containing anti-CTLA-4 antibody (see Example 1) at 37°C for 1 hour. The plate was washed and incubated with horseradish peroxidase (HRP)-conjugated goat anti-human IgG (H+L) antibody (A18805, Life technologies) at 37°C for 1 hour. 100 μL of tetramethylbenzidine (TMB) was added, and the plate was incubated at room temperature for 15 minutes. 50 μL of 1N HCl was added to terminate the reaction, and OD450 nm was measured using an ELISA plate reader.

[0226] Meanwhile, the 293-hCTLA-4 cells prepared in step 1 were cultured and used to measure the antibody binding activity. The cells were treated with an enzyme-free cell dissociation solution (Versene solution, Invitrogen) and then collected. BSA was added to the cell suspension to a final concentration of 1%, and the cells were blocked on ice for 30 minutes and then washed twice with HBSS. The cells were collected after centrifugation and eluted at 2 x 10 6Cells / mL were resuspended in FACS buffer (HBSS+1% BSA, v / v). 100 μL of the cell suspension was then added to each well of a 96-well plate. 100 μL of 293F cell culture medium containing anti-CTLA-4 antibodies (see Example 1) was added to each well of a 96-well plate and incubated on ice for 1 hour. The cells were washed twice with FACS buffer, and 100 μL of Alexa 488-labeled anti-human (H+L) antibody (Invitrogen) was added to a 96-well plate and incubated on ice for 1 hour. The sample was washed 3 times with FACS buffer, and 100 μL of fixative buffer (4% paraformaldehyde v / v) was added to each well and incubated for 10 minutes. The cells were then washed twice with FACS buffer and resuspended in 100 μL of FACS buffer. The mean fluorescence intensity (MFI) was measured using FACSCalibur (BD).

[0227] Step 3: Production and purification of lead candidate antibodies

[0228] The antibody concentration from HEK293 cells is approximately 1-10 μg / mL and varies greatly. In addition, FBS and culture medium components may interfere with analysis. Therefore, it is necessary to produce and purify antibodies on a small scale (1-5 mg).

[0229] A construct containing a nucleotide sequence encoding an anti-CTLA-4 antibody (as listed in Table 1) was introduced into 293 cells. 6-7 days after transfection, the supernatant containing the target antibody was harvested by centrifugation and filtration. The monoclonal antibody was purified by passing it through a 2 mL protein G column (GE Healthcare). The protein G column was first equilibrated with PBS buffer (pH 7.2), and then the hybridoma culture supernatant was applied to the equilibrated protein G column at a constant flow rate of 3 mL / min. Each column was then washed with PBS buffer having a volume 3 times greater than the volume of the column. The anti-CTLA-4 antibody was then eluted with elution buffer (0.1 M acetate buffer, pH 2.5), and the UV absorbance of the eluate was monitored using a UV detector (A280 UV absorption peak). 10% 1.0 M Tris-HCl buffer was added to the eluate to neutralize the pH, and the sample was sterile filtered by passing it through a 0.22 micron filter. Sterile-filtered purified anti-CTLA-4 antibody was obtained.

[0230] The concentration of the purified anti-CTLA-4 antibody was determined by UV absorbance (A280 / 1.4), and the purity and endotoxin levels were measured (Lonza kit). The purified anti-CTLA-4 antibody had an endotoxin concentration of less than 1.0 EU / mg.

[0231] Example 3 - Characterization of lead candidate antibodies

[0232] 293 cells were stably transfected with a pTT5 plasmid containing a nucleic acid sequence encoding human CTLA-4 (SEQ ID NO: 196) to generate 293F cells stably expressing human CTLA-4 (referred to herein as 293-hCTLA-4 cells). Additional 293 cells were stably transfected with a pIRES plasmid containing a nucleic acid sequence encoding full-length cynomolgus monkey CTLA-4 (SEQ ID NO: 198) to generate 293 cells stably expressing cynomolgus monkey CTLA-4 (referred to herein as 293-cynoCTLA-4 cells). 293-hCTLA-4 and 293-cynoCTLA-4 cells were cultured and expanded to 90% confluence in a T-75 culture flask. The culture medium was aspirated and the cells were washed twice with HBSS (Hanks balanced salt solution, Invitrogen). The cells were treated with an enzyme-free cell dissociation solution (Versene solution, Invitrogen) and collected. The cells were then washed twice with HBSS, the cell count was determined, and 2 x 10 6 Resuspend the cells in HBSS at 10 cells / mL. Add BSA to the cell suspension to a final concentration of 1%, and block the cells on ice for 30 minutes, then wash twice with HBSS. Collect the cells after centrifugation and use 2x 10 6 Cells / mL were resuspended in FACS buffer (HBSS+1% BSA, v / v). 100 μL of the cell suspension was then added to each well of a 96-well plate. 100 μL of purified anti-CTLA-4 antibody or control antibody from Example 2 was added to each well of a 96-well plate and incubated on ice for 1 hour, wherein the heavy and light chains of the ipilimumab analogs have the amino acid sequences of SEQ ID NO: 199 and SEQ ID NO: 200, respectively. The cells were washed twice with FACS buffer, and 100 μL of Alexa 488-labeled anti-human (H+L) antibody (Invitrogen) was added to a 96-well plate and incubated on ice for 1 hour. The sample was washed 3 times with FACS buffer, and 100 μL of fixation buffer (4% paraformaldehyde, v / v) was added to each well and incubated for 10 minutes. The cells were then washed twice with FACS buffer and resuspended in 100 μL of FACS buffer.

[0233] The mean fluorescence intensity (MFI) was determined using FACS Calibur (BD), and the results are shown in Figure 1 and Figure 2 The antibody from Example 2 had comparable binding activity to human or cynomolgus monkey CTLA as the ipilimumab analog on 293F cells.

[0234] Example 4 - Determination of the ability of anti-CTLA-4 antibodies to block CTLA-4 binding to B7.1

[0235] A cell-based receptor-ligand binding assay was performed to determine the ability of anti-CTLA-4 antibodies to block the binding of CTLA-4 to its ligand B7.1. Recombinant B7.1 was stained with EZ-LINK NHS-PEG12-biotin (ThermoScientific #21312) according to the manufacturer's instructions. ECD -Fc protein (B71-H5259, Acro Bio) was biotinylated. Concentrated biotinylated B7.1 ECD -Fc protein, and free label was removed using Amicon centrifugal filters (10 kDa cutoff). The extracellular domain of B7.1 corresponds to amino acids Val35-Asn242 of protein P33681 in the Uniprot database.

[0236] The 293-hCTLA-4 cells prepared in Example 3 were cultured and expanded to 60-80% confluence in a T-75 culture flask. The culture medium was aspirated and the cells were washed twice with PBS. The cells were treated with an enzyme-free cell dissociation solution (Versene solution, Invitrogen) and collected. The dissociation solution was neutralized by adding 8 mL of culture medium and the cell count was determined. The cells were centrifuged at 300 g for 5 minutes and then centrifuged at 1 x 10 6 Resuspend cells / mL in blocking buffer (PBS buffer containing 2% BSA, pH 7.4, w / v). Block the cells at 37°C for 15 minutes. Simultaneously, block the wells of a 96-well round-bottom plate with 200 μL of blocking buffer at 37°C for 1 hour. Discard the blocking buffer and dispense 200 μL of cells into each well of the 96-well plate (2 x 10 5 The plate was centrifuged at 500 g for 5 minutes and the supernatant was discarded. The cells were resuspended in 100 μL of anti-CTLA-4 antibody prepared in blocking buffer with different concentrations. 100 μL of biotinylated B7.1 ECD -Fc (60 μg / mL in blocking buffer) was added to each well of a 96-well plate and mixed by gentle shaking. The plate was incubated at 4°C for 90 minutes and washed twice with 200 μL of blocking buffer. The blocking buffer was discarded and the cells were resuspended in 100 μL of streptavidin-Alexa 488 solution (Invitrogen, 1:500 in blocking buffer) and incubated at 4°C for 1 hour. The plate was washed three times with blocking buffer and 200 μL of blocking buffer was added.

[0237] The mean fluorescence intensity (MFI) was measured using FACS Calibur (BD). Figure 3As shown in , the results demonstrate that anti-CTLA-4 antibodies can block the binding of cell-expressed CTLA-4 to its ligand B7.1 at a level comparable to that of the ipilimumab analog.

[0238] Example 5 - Anti-CLTA-4 HCAb antibody promotes IL-20 release

[0239] Step 1 PBMC stimulation test

[0240] 100 μL of PBMC (containing 1x 10 5 Cells were added to the wells of a 96-well plate. 50 μL of each test antibody at various concentrations was then added to the 96-well plate and incubated at room temperature for 15 minutes. 50 μL of 100 ng / mL SEB was added to each well and incubated at 37°C, 5% CO2 for 72 hours. The supernatant was collected and stored at -20°C until analysis.

[0241] Step 2: Detection of interleukin IL-2 secretion by ELISA

[0242] Following the operating instructions provided by the manufacturer, the level of IL-2 in the culture supernatant was quantified using human IL-2Quantikine ELISA kit (DY202, R&D Systems). In brief, anti-IL-2 polyclonal antibodies were coated on ELISA microplates, and 100 μL of culture supernatant and standard were added to each well and incubated at room temperature for 2 hours. The plate was washed 4 times with wash buffer, followed by addition of HRP-conjugated anti-human IL-2 antibodies, and incubated at room temperature for 2 hours. After washing, chromogenic substrate (5120-0077, SeraCare) was added, and incubated for 30 minutes at room temperature in the dark, and the reaction was terminated by adding stop solution (E661006-0200, BBI Life sciences).

[0243] The absorbance at 450 nm was measured using an ELISA plate reader and the Figure 4 As shown in Figures A and 4B, the results demonstrate that some anti-CTLA-4 antibodies can increase IL-2 secretion at low concentrations compared to human IgG (AB170090, Crown Bio) or ipilimumab analogs.

[0244] Example 6 - Anti-CTLA-4 HCAb antibody binds to human CTLA-4 but not to murine CTLA-4 in an ELISA assay

[0245] Recombinant human or mouse CTLA4-his protein (CT4-H5229 for human protein and CT4-M52H5 for mouse protein, Acro Bio) was diluted to a concentration of 2 μg / mL in PBS, and 100 μL of diluted CTLA-4-his protein was added to each well of an ELISA microplate, which was incubated overnight at 4°C to coat the plate with the recombinant protein. The plate was then blocked with ELISA blocking solution (containing 2% BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer, w / v) at 37°C for 2 hours, and then incubated with anti-CTLA-4 antibody at various concentrations for 1 hour at 37°C. The plate was washed and incubated with horseradish peroxidase (HRP)-conjugated goat anti-human IgG (H+L) antibody (A18805, Life technologies) at 37°C for 1 hour. 100 μL of tetramethylbenzidine (TMB) was added, and the plate was incubated at room temperature for 15 minutes. 50 μL of 1N HCl was added to terminate the reaction, and OD450 nm was measured using an ELISA plate reader.

[0246] The data shows Figure 5 A and Figure 5 In B, all clones were shown to bind to human CTLA-4 but to little to murine CTLA-4.

[0247] Example 7 - Anti-CTLA-4 antibody mutants promote IL-20 release

[0248] Some of the above antibodies and their mutants were subjected to T cell stimulation assays to examine the effects of these antibodies on T cell stimulation by blocking the binding of CTLA-4 to its ligands B7.1 and B7.2.

[0249] Antibody mutants were prepared by PCR using S239D and I332E mutations in the Fc constant domain of HCAb clone 5, clone 11, clone 22, clone 25, and clone 30. The mutated antibodies were expressed in HEK293 cells and purified as described in Example 2. The nucleic acid and amino acid sequences of the antibody mutants were determined using standard molecular biology methods and are summarized in Table 2.

[0250] Table 2. Nucleic acid and amino acid sequences of anti-CTLA-4 antibody mutants

[0251]

[0252] na: nucleic acid; aa: amino acid

[0253] PBMC stimulation assays and detection of interleukin IL-2 secretion by ELISA were performed as described in Example 5.

[0254] Figure 6 The results in A and 6B demonstrate that the anti-CTLA-4 antibody mutants promoted greater IL-2 secretion when compared to a human IgG1 isotype control and its parental clone.

[0255] Example 8 - PTM-removed anti-CTLA-4 antibody variants

[0256] The amino acid sequences of seven HCAb clones (CL22, CL25, CL5, CL3, CL11, CL30, and CL24) were aligned with the gene IGHV3-53*01 and are shown in Table 3. Differences from the germline gene and PTM sites are highlighted. Sequences are numbered using the Chothia numbering scheme.

[0257] The germline gene IGHV3-53 does not naturally have an N-glycosylation motif, and those N-glycosylation motifs carried in the 7 HCAb clones are formed by somatic mutation. Therefore, one method of removing this motif is to replace it with the corresponding residue in the germline, for example, replacing the NVS motif in the CDR1 of CL11 with the TVS of the germline. Based on the concept of CDR transplantation for humanization, an alternative method is explored by combining PTM removal with "germlining". In the second method, the CDR of each HCAb is transplanted into the germline IGHV3-53 framework, and the key framework residues from the parent HCAb are also retained. For some antibodies, amino acid mutations in specific residues that may affect the binding activity between CTLA-4 and the antibody are restored. The nucleic acid and amino acid sequences of human anti-CTLA-4 HCAb antibody variants with PTM removed are listed in Table 4.

[0258] Table 3. Differences in HCAb amino acid sequences compared to germline sequences

[0259]

[0260]

[0261] Table 4. Nucleic acid and amino acid sequences of PTM-removed anti-CTLA-4 antibody variants

[0262]

[0263]

[0264] na: nucleic acid; aa: amino acid

[0265] Example 9 - Cell-based binding activity of PTM-removed anti-CTLA-4 antibody variants

[0266] A cell-based binding assay was performed to determine the ability of the PTM-removed anti-CTLA-4 antibody variants to bind to human CTLA-4. The assay procedure was similar to that described in Example 3. Briefly, 293F-hCTLA-4 cells were harvested using an enzyme-free cell dissociation solution (Versene solution, Invitrogen), then neutralized with culture medium, and the cell count was determined. The cells were centrifuged and incubated at 37°C for 1 x 10 6 10 cells / mL were blocked in blocking buffer (PBS buffer containing 2% BSA, pH 7.4, w / v) for 15 minutes. 200 μL of cells were dispensed into each well of a 96-well plate (2 x 10 5 cells / well). The plate was centrifuged and the supernatant discarded. The cells were resuspended in 100 μL of anti-CTLA-4 antibody prepared in blocking buffer. The plate was incubated at 4°C for 90 minutes and washed twice. After discarding the blocking buffer, the cells were resuspended in 100 μL of Alexa 488-labeled anti-human (H+L) antibody (1:500, Invitrogen) and incubated at 4°C for 1 hour. The cells were washed and resuspended in 200 μL of blocking buffer. The mean fluorescence intensity (MFI) was measured using a FACS Calibur (BD).

[0267] like Figure 7 As shown in , the results demonstrate that PTM-depleted anti-CTLA-4 antibody variants bind to cell-expressed human CTLA-4.

[0268] Example 10 - Blocking activity of PTM-removed anti-CTLA-4 antibody variants on CTLA-4-B7.1 and B7.2 interactions

[0269] Cell-based receptor ligand binding assays were performed as described in Example 4 to determine the ability of PTM-removed anti-CTLA-4 antibody variants to block binding of CTLA-4 to its ligand B7.1.

[0270] like Figure 8 As shown in , the results demonstrated that the PTM-depleted anti-CTLA-4 antibody variant blocked the binding of cell-expressed CTLA-4 to its ligands B7.1 and B7.2 at levels comparable to those of the ipilimumab analog (data not shown).

[0271] Example 11 - PTM-removed anti-CTLA-4 antibody variants promote IL-20 release

[0272] PBMC stimulation and quantification of IL-20 levels were performed as described in Example 5.

[0273] like Figure 9As shown in , the results demonstrate that anti-CTLA-4 antibody variants with PTM removal can still promote IL-2 secretion. Anti-CTLA-4 antibodies with S239D and I332E mutations and PTM removal induced increased IL-2 secretion compared to anti-CTLA-4 antibodies with S239D and I332E mutations but without PTM removal.

[0274] Example 12 - Binding affinity and dissociation constants of PTM-removed anti-CTLA-4 antibody variants

[0275] The dissociation constant was determined by Biacore T200 (GE Healthcare) following the manufacturer's instructions for the instrument. Briefly, an anti-CTLA-4 antibody diluted at 1 μg / mL in 10 mM NaOAc (pH 5.0, Sigma) was immobilized on a flow cell of a SCM5 series sensor chip. The remaining active ester groups were blocked with 1 M ethanolamine (pH 8.5). Using HBS-EP+ as running buffer, recombinant human CTLA-4-his (CT4-H5229, AcroBio) and cynomolgus monkey CTLA-4-his protein (CT4-C5227, AcroBio) with five serial dilution concentrations were injected into the flow cell at 30 μL / min, with an association time of 180 s. The buffer flow was maintained for 600 s to dissociate. The KD value for each interaction between the antibody and antigen was evaluated using Biacore T200 evaluation software 1.0 and a 1:1 combined fitting model.

[0276] The results are shown in Figure 10 and Table 5. The binding affinities of both clones were similar to that of the ipilimumab analogs.

[0277] Table 5. Human anti-CTLA-4 antibodies and human CTLA-4 as determined by Biacore T200 ECD -his protein and cynomolgus monkey CTLA-4 ECD Binding kinetics and affinity of -his proteins

[0278] Clone ID protein KD(M) ka(1 / Ms) kd(1 / s) CL5'-dPTM' <![CDATA[Human CTLA-4 ECD -his]]> 4.28E-11 5.35E+06 2.29E-04 CL5'-dPTM' <![CDATA[Cynomolgus monkey CTLA-4 ECD -his]]> 5.91E-11 5.21E+06 3.08E-04 CL5'-eA-dPTM' <![CDATA[Human CTLA-4 ECD -his]]> 1.40E-11 5.40E+06 7.58E-05 CL5'-eA-dPTM' <![CDATA[Cynomolgus monkey CTLA-4 ECD -his]]> 2.43E-11 4.55E+06 1.10E-04 Ipilimumab analogs <![CDATA[Human CTLA-4 ECD -his]]> 7.32E-11 1.23E+06 8.98E-05 Ipilimumab analogs <![CDATA[Cynomolgus CTLA-4 ECD -his]]> 3.47E-10 3.73E+06 1.29E-03

[0279] Example 13 - In vitro ADCC functional analysis

[0280] To confirm the putative NK-dependent cytotoxic activity of human anti-CTLA-4 antibodies, antibody-dependent cell-mediated cytotoxicity (ADCC) assays were performed on both CTLA-4-expressing CHO-K1 cells and in vitro stimulated Treg cells expressing CTLA-4.

[0281] As described in step 1 of Example 2, CHO-K1 cells expressing CTLA-4 were adjusted to 2×10 cells / mL using ADCC medium (RPMI 1640 without phenol red, 10% FBS, and 1% penicillin / streptomycin). 5 50 μL of cell suspension (1×10 4 100 μL of NK92 cells stably transfected with FcγRⅢ158V were adjusted with ADCC medium (without phenol red) so that the ratio of effector to target cells was 5: 1. The plate was then incubated at 37°C for 6 hours. After 6 hours of incubation, the plate was centrifuged and 50 μL of each supernatant was transferred to a new plate. The supernatant was incubated with 50 μL of LDH detection buffer at room temperature for 30 minutes, and the absorbance at 490 nm was measured. For the maximum cell lysis control, 50 μL of CHO-K1 cells expressing CTLA-4, 50 μL of ADCC medium, and 100 μL of 1% triton-X100 buffer were added for LDH detection. For the minimum cell lysis control, 50 μL of CHO-K1 cells expressing CTLA-4 and 150 μL of ADCC medium were added for LDH detection. The absorbance at 492 / 650 nm was measured. The percentage of cell lysis was calculated as 100*(absorbance of sample – absorbance of background) / (absorbance of maximum release – absorbance of minimum release). The percentage of all cell lysis values ​​was calculated using GraphPad Prism 5.0.

[0282] like Figure 11 As shown in , the results demonstrated that the clone CL5-dPTM' antibody induced ADCC against CTLA-4-expressing CHO-K1 cells, and that the clone CL5-eA-dTPM' with additional S239D and I332E mutations showed higher ADCC activity than the ipilimumab analog.

[0283] The Treg cells expressing in vitro stimulation of CTLA-4 are derived from the initial CD4+T cells isolated in vitro. First, initial CD4+T cells were isolated from primary PBMCs according to the manufacturer's instructions (Miltenyi, 130-094-131). Then, initial CD4+T cells were activated by incubation for three days with Dynabeads human T activator CD3 / CD28 (1:1) (Thermo, 11131D), 10ng / ml IL-2 (PeproTech, 200-02-B) and 20ng / ml TGF-β1 (PeproTech, 100-21). On the day of the ADCC killing experiment, the stimulated Treg cells were adjusted to 1×10 with ADCC culture medium (containing RPMI 1640, 10% FBS and 1% penicillin / streptomycin without phenol red). 6 1×10 cells / mL was added with 5 μl of Calcein AM (Therom, C34851, stock solution was prepared as 50 μg per 50 μl of DMSO). 6 The cells were stained at 37°C for 1 hour. Treg cells were washed 3 times with ADCC medium. 50 μL of Treg cell suspension (5×10 3 10 viable cells) were added to each well of a V-bottom 96-well plate. The test antibody was serially diluted in ADCC medium (without phenol red), and 50 μL of each resulting solution was added to the wells in triplicate. The final antibody concentrations were: 1 pM, 10 pM, 100 pM, 1 nM, 10 nM, and 100 nM. The plate was incubated at room temperature for 30 minutes. Fresh PBMC (Miaotong) was adjusted to 5 × 10 cells / well with ADCC medium (without phenol red). 6 cells / mL. By adding 50 μL of fresh PBMCs to the stained Treg cells, the effector to target cell ratio was 50:1. The plate was then incubated at 37°C for 2 hours. After a 2-hour incubation, the plate was centrifuged and 100 μL of each supernatant was transferred to a new plate. The supernatant was measured using the Enspire instrument. For the maximum cell lysis control, 50 μL of Calcein AM-stained Treg cells, 50 μL of ADCC medium, and 100 μL of 1% triton-X100 buffer were added for detection of released Calcein AM. For the minimum cell lysis control, 50 μL of Calcein AM-stained cells and 150 μL of ADCC medium were added for detection of released Calcein AM. The absorbance at 520 / 650 nm was measured. The percentage of specific killing was calculated as 100*(absorbance of sample – absorbance of background) / (absorbance of maximum release – absorbance of minimum release). The percentage of all specific killing values ​​was calculated using GraphPad Prism 5.0.

[0284] The results showed that human anti-CTLA-4 antibodies induced ADCC against CTLA-4-expressing Treg cells, and antibodies with S239D and I332E mutations exhibited higher ADCC activity than non-mutated antibodies.

[0285] Example 14 - Pharmacokinetic Study of Anti-CTLA-4 Antibodies

[0286] Step 1: Single-dose anti-CTLA-4 antibody treatment in mice

[0287] Serum concentrations of anti-CTLA-4 antibodies were measured by injecting male C57BL / 6 mice with 3 mg / kg of anti-CTLA-4 antibody via the tail vein. Animals were manually restrained, and approximately 100 μL of blood was collected via retro-orbital puncture at time points: pre-dose, 0.167, 1, 4, 8, 24 hours, and 2, 4, 7, and 14 days after antibody injection. Final collection was performed via cardiac puncture. Blood samples were placed at room temperature and centrifuged at 2,000 × g for 5 minutes at 4°C to obtain serum samples.

[0288] Step 2: Serum concentration of anti-CTLA-4 antibodies measured by ELISA

[0289] All serum samples were first diluted 20-fold in assay diluent. Additional dilutions were performed in 5% mouse serum (PBS, v / v). Recombinant human CTLA4-his protein (Acro Bio) was diluted to a concentration of 0.5 μg / mL in PBS, and 50 μL of diluted CTLA-4-his protein sample was added to each well of an ELISA microplate, which was incubated overnight at 4°C to coat the plate with the recombinant protein. The plate was then blocked with ELISA blocking solution (containing 2% BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer, w / v) at 37°C for 2 hours. The blocking buffer was aspirated, and the plate was incubated with the diluted serum sample at 37°C for 1 hour. The plate was washed three times with wash buffer (PBS + 0.01% (v / v) Tween 20) and incubated with horseradish peroxidase (HRP)-conjugated goat anti-human IgG (Fc) antibody (A0170, Sigma) at 37°C for one hour. 100 μL of tetramethylbenzidine (TMB) was added, and the plate was incubated at room temperature for 15 minutes. 100 μL of 0.1N HCl was added to each well to terminate the reaction. The absorbance at 450 nm was measured using an ELISA reader (SpectraMax M2).

[0290] The corresponding serum concentrations of anti-CTLA-4 antibodies are shown in Figure 12 The detailed data are listed in Table 6.

[0291] Table 6. Mean serum concentrations of antibodies in mice following IV administration at 3 mg / kg

[0292]

[0293]

[0294] In addition, the tumor-to-serum ratio of anti-CTLA4 HCAb concentrations was measured in C57BL / 6 mice bearing MC38 tumors. Mice were injected with 3 mg / kg of anti-CTLA-4 antibody via the tail vein to measure serum and tumor-resident concentrations of anti-CTLA-4 antibody. Animals were manually restrained and approximately 100 μL of blood / time point was collected via retro-orbital puncture, with the time points being 8 and 24 hours after injection. Final collection was performed via cardiac puncture. Blood samples were kept at room temperature and centrifuged at 2,000×g for 5 minutes at 4°C to obtain serum samples. As in step 2, serum and tumor concentrations of anti-CTLA-4 antibodies were also tested by ELISA.

[0295] like Figure 13 As shown in Figure 3, the tumor-to-serum ratio of anti-CTLA4 HCAb concentration in the CL5-eA-dPTM' group was approximately double that in the ipilimumab analog group, likely due to the unique characteristics of heavy chain-only HCAb antibodies. This higher tumor-to-serum distribution may result in higher tumor tissue penetration of HCAb antibodies.

[0296] Example 15 - Better survival of MC38 tumor-bearing mice using human anti-CTLA-4 antibodies

[0297] Cryopreserved murine colon cancer MC-38 cell line was recovered and cultured at 37°C in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin to obtain sufficient cells for tumor implantation. The cultured MC-38 cells were harvested and plated at 1×10 7 The cells were resuspended in PBS at a density of 10 cells / ml with a viability of >90% and implanted subcutaneously into the right flank of 120 hCTLA-4 knock-in mice (GempharmaTech). Five days after tumor inoculation, 81 mice with tumors ranging from 26 to 64 mm in size were selected. 3 (The average tumor size was 40 mm 3) mice were randomly divided into 9 groups based on their tumor volume using stratified randomization, with 9 mice in each group. Treatment started on the day of randomization (defined as D0). Group 1 was treated with hIgG1 at 10 mpk on D0, D3, D6, D10, D13, and D16. ip treatment; Group 2 was treated with CL20'-eA (sequence in Table 7) ip at 5.4 mpk on D0, D3, D6, D10, D13, D16; Group 3 was treated with ipilimumab analogue ip at 10 mpk on D0, D3, D6, D10, D13, D16; Group 4 was treated with ipilimumab analogue ip at 1 mpk on D0, D3, D6, D10, D13, D16; Group 5 was treated with CL5'-dPTM' i.p. at 5.4 mpk on D0, D3, D6, D10, D13, D16; Group 6 was treated with ipilimumab analogue ip at 1 mpk on D0, D3, D6, D10, D13, D16; Group 1 was treated with CL5'-dPTM'i.p. at 0.54 mpk on D0, D3, D6, D10, D13, and D16; Group 7 was treated with CL5'-eA-dPTM'i.p. at 5.4 mpk on D0, D3, D6, D10, D13, and D16; Group 8 was treated with CL5'-eA-dPTM'i.p. at 1.5 mpk on D0, D3, D6, D10, D13, and D16; and Group 9 was treated with CL5'-eA-dPTM'i.p. at 0.54 mpk on D0, D3, D6, D10, D13, and D16;.

[0298] Table 7. Nucleic acid and amino acid sequences of clone CL20'-eA

[0299]

[0300] na: nucleic acid; aa: amino acid

[0301] Tumor size was measured three times a week during the treatment period. 3 ) were euthanized. The time from the start of treatment to the end of treatment was considered as their survival time. Survival curves were drawn using the Kaplan-Meier method. The median survival time (MST) of each group was calculated. The life expectancy (ILS) was calculated according to the following formula:

[0302] ILS(%)=(MSTT 治疗 -MST 媒介物 ) / MST 媒介物 x 100%).

[0303] According to the National Cancer Institute criteria, an ILS (%) > 25% would be considered a biologically significant survival benefit.

[0304] The relative change in body weight (RCBW) of each mouse was calculated according to the following formula: RCBW (%) = (BW i –BW0) / BW0×100%, where BW i refers to the mean body weight on day i, and BW0 refers to the mean body weight on day 0.

[0305] Tumor volume (TV) was calculated based on the following formula: tumor volume = (length × width 2 ) / 2.

[0306] The tumor growth inhibition rate (TGI%) of each drug-treated group was calculated according to the following formula: TGI% = [1-TV i / TV vi ]*100%, of which TV i refers to the average tumor volume of the drug-treated group on the first day, and TV vi Refers to the mean tumor volume of the vehicle group on day i.

[0307] The mean and standard error of the mean (SEM) of mouse body weight, RCBW, and tumor volume for each group were calculated using Microsoft Excel 2007. GraphPad Prism 5 was used to plot body weight, relative change in body weight, tumor growth curves, and tumor growth inhibition. Tumor growth between groups was analyzed using a two-way RM ANOVA. Kaplan-Meier survival curves were analyzed using the Log-Rank test. A P value of < 0.05 was considered statistically significant.

[0308] The entire study was terminated at D65. The individual tumor growth curves of each group are shown in Figure 14 A. Kaplan-Meier survival curves of animals from time to endpoint are shown in Figure 14 B. Figure 14 C, all treatments were tolerated without any adverse effects. In most of these groups, tumor volumes reached 2000 mm 3 In Group 7, 3 of 9 mice treated with CL5'-eA-dPTM' at 5.4 mpk were tumor-free on D65. In Group 9, 2 of 9 mice treated with CL5'-eA-dPTM' at 5.4 mpk were tumor-free on D65.

[0309] The MST was calculated for each group and is shown in Table 8. The MST for both the vehicle-treated hIgG1 (5.4 mpk) and CL20'-eA (5.4 mpk) groups was 14 days. The MSTs for the treatment groups treated with ipilimumab analogs at 10 mpk and 1 mpk, CL5'-dPTM' at 5.4 mpk and 0.54 mpk, and CL5'-eA-dPTM' at 5.4 mpk, 1.5 mpk, and 0.54 mpk were 14, 23, 19, 21, 14, 40, 21, and 28 days, respectively. As can be seen in Table 8, compared to the vehicle-treated hIgG1 group, the ILS in the treatment groups was 64.3%, 35.7%, 50%, 0%, 185.7%, 50%, and 100%. The median survival time was significantly increased by 10 mpk of the ipilimumab analog, 5.4 mpk of CL5'-dPTM', and 5.4 mpk, 1.5 mpk, and 0.54 mpk of CL5'-eA-dPTM'. Mice treated with 5.4 mpk of CL5'-eA-dPTM' exhibited better ILS than mice treated with 10 mpk of the ipilimumab analog.

[0310] Table 8. Survival analysis

[0311]

[0312] Note: ILS = (MST Gn -MST G1 ) / MST G1 *100%; P values ​​are for all groups compared with the G1 group.

[0313] Example 16 - Anti-CTLA-4 HCAb inhibits MC38 tumor growth in hCTLA-4 knock-in mice

[0314] Cryopreserved murine colon cancer MC-38 cell line was recovered and cultured at 37°C in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin to obtain sufficient cells for tumor implantation. The cultured MC-38 cells were harvested and plated at 5×10 6 The cells were resuspended in PBS at a density of 10 cells / ml with a viability of >90% and implanted subcutaneously into the right flank of 60 hCTLA-4 knock-in mice. Seven days after tumor inoculation, 30 mice with a tumor size of 102 mm were selected. 3Tumor-bearing mice with an average tumor size of 100 mg / kg were randomly divided into 5 groups (n=6) based on their tumor size. Treatment began on the day of randomization (defined as D0). Mice enrolled were treated with hIgG1 (0.5 mpk), ipilimumab analogs (0.5 and 0.2 mpk) and CL5'-eA-dPTM' (0.27 and 0.1 mpk) intraperitoneally (ip) on D0, D6, D9, D13, D16 and D19, respectively. Mice were monitored daily and body weights were recorded on weekdays. Tumor size was measured twice a week during treatment.

[0315] Relative change in body weight (RCBW), tumor volume (TV), and tumor growth inhibition (TGI%) were calculated as in Example 16. The mean and standard error of the mean (SEM) of mouse body weight, RCBW, and tumor volume were calculated for each group using Microsoft Excel 2007. Body weight, relative change in body weight, tumor growth curves, and tumor growth inhibition were plotted using GraphPad Prism 5. Tumor growth between groups was analyzed using a two-way RM ANOVA. Kaplan-Meier survival curves were analyzed using the Log-Rank test. A P value of <0.05 was considered statistically significant.

[0316] Tumor growth curves are shown in Figure 15 A. Figure 15 As observed in Figure 2B, all treatments were tolerated without any adverse effects. Mice in the CL5'-eA-dPTM' (0.1 mpk) and 0.27 mpk) and ipilimumab (0.5 mpk) analogue groups showed significant tumor growth inhibition from D20 to D30 compared to mice in the hIgG1 (0.2 mpk) group. Meanwhile, ipilimumab analogue treatment at 0.2 mg / kg did not show significant tumor growth inhibition compared to hIgG1 (0.2 mpk). At the same dose, CL5'-eA-dPTM' promoted better tumor growth inhibition than ipilimumab analogue.

[0317] While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope of the invention. Sequence Listing <110> Hebo Pharmaceuticals (Shanghai) Co., Ltd. <120> Antibodies that bind to CTLA-4 and their uses <130> 55543 00001 <150> US62 / 607917 <151> 2017-12-20 <160> 200 <170> PatentIn version 3.5 <210> 1 <211> 1113 <212> DNA <213> Homo sapiens <400> 1 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gattggagga ggcgagatcc agccgggggg gtccctgaga 120 ctctcgtgtg cagcctctgg gttcaacgtc agtaagaact acatgagttg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtaatctata gtggtggtgg tacagaatac 240 gcagcctccg tgaagggccg atttaacatc tccagagaca gttccaagaa tatgttgtat 300 cttcaaatga acaacctgag agccgaggac acggccgtgt attattgtgc gagggccgtc 360 cccaggcccc atgggactga tatctggggc caagggacaa tggtcaccgt ctcttcagag 420 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 480 ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 540 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 600 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 660 aacagcacgt accgggtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 720 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc 780 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 acgcagaaga gcctctccct gtctccgggt aaa 1113 <210> 2 <211> 371 <212> PRT <213> Homo sapiens <400> 2 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ile Gly Gly Gly Glu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Asn Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Gly Thr Glu Tyr 65 70 75 80 Ala Ala Ser Val Lys Gly Arg Phe Asn Ile Ser Arg Asp Ser Ser Lys 85 90 95 Asn Met Leu Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro Arg Pro His Gly Thr Asp Ile 115 120 125 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser Cys 130 135 140 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 145 150 155 160 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 165 170 175 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 180 185 190 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 195 200 205 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 210 215 220 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 225 230 235 240 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 245 250 255 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 260 265 270 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 275 280 285 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 290 295 300 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 305 310 315 320 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 325 330 335 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 340 345 350 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 355 360 365 Pro Gly Lys 370 <210> 3 <211> 118 <212> PRT <213> Homo sapiens <400> 3 Glu Val Gln Leu Val Glu Ile Gly Gly Gly Glu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Gly Thr Glu Tyr Ala Ala Ser Val Lys 50 55 60 Gly Arg Phe Asn Ile Ser Arg Asp Ser Ser Lys Asn Met Leu Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Arg Pro His Gly Thr Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 <210> 4 <211> 10 <212> PRT <213> Homo sapiens <400> 4 Gly Phe Asn Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 5 <211> 16 <212> PRT <213> Homo sapiens <400> 5 Val Ile Tyr Ser Gly Gly Gly Thr Glu Tyr Ala Ala Ser Val Lys Gly 1 5 10 15 <210> 6 <211> 10 <212> PRT <213> Homo sapiens <400> 6 Ala Val Pro Arg Pro His Gly Thr Asp Ile 1 5 10 <210> 7 <211> 1116 <212> DNA <213> Homo sapiens <400> 7 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggcgga gactggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaatgtc agtaagaatt acatgagctg ggtccgccag 180 gttccaggga aggggctgga gtgggtcgca gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcatcctc tccagagaca attccgacaa cacactgtat 300 cttcaaatga acaacctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 8 <211> 372 <212> PRT <213> Homo sapiens <400> 8 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Asn Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 9 <211> 119 <212> PRT <213> Homo sapiens <400> 9 Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Asn Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 10 <211> 10 <212> PRT <213> Homo sapiens <400> 10 Gly Phe Asn Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 11 <211> 16 <212> PRT <213> Homo sapiens <400> 11 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 12 <211> 11 <212> PRT <213> Homo sapiens <400> 12 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 13 <211> 1116 <212> DNA <213> Homo sapiens <400> 13 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggagga gactggagga gacttgatcc agccgggggg gtccctgaga 120 ctctcctgta cagcctctgg gtttaacgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaggggccg atttaccatc tccagagaca gttccacgaa cacggtgttt 300 cttcaaatga acagcctgag agccgaggac tcggccgtgt attactgtgc gagagccgtc 360 ccaagtgccc ctagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 14 <211> 372 <212> PRT <213> Homo sapiens <400> 14 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe 35 40 45 Asn Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr 85 90 95 Asn Thr Val Phe Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 15 <211> 119 <212> PRT <213> Homo sapiens <400> 15 Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Asn Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr Asn Thr Val Phe Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 16 <211> 10 <212> PRT <213> Homo sapiens <400> 16 Gly Phe Asn Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 17 <211> 16 <212> PRT <213> Homo sapiens <400> 17 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 18 <211> 11 <212> PRT <213> Homo sapiens <400> 18 Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 1 5 10 <210> 19 <211> 1119 <212> DNA <213> Homo sapiens <400> 19 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gtccggggga ggcttagttc agcctggggg gtccctgaga 120 ctctcctgtg cagcctctgg attcaccttc agtaactact ggatgcactg ggtccgccaa 180 gctccaggga aggggctggt gtgggtctca cgtattaata gtgatgggag taacacaacc 240 tacgcggact ccgtgaaggg ccgattcacc atctccagag acaacgccaa gaacacgctg 300 tatctgcaaa tgaacagtct gagagccgag gacacggctg tgtattactg tgcaagatgg 360 ttcggggagc actactacgg aatggacgtc tggggccaag ggaccacggt caccgtctct 420 tcagagccca aatcttgtga caaaactcac acatgcccac cgtgcccagc acctgaactc 480 ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc 540 cggacccctg aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag 600 ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc gcgggaggag 660 cagtacaaca gcacgtaccg ggtggtcagc gtcctcaccg tcctgcacca ggactggctg 720 aatggcaagg agtacaagtg caaggtctcc aacaaagccc tcccagcccc catcgagaaa 780 accatctcca aagccaaagg gcagccccga gaaccacagg tgtacaccct gcccccatcc 840 cgggaggaga tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc 900 agcgacatcg ccgtggagtg ggagagcaat gggcagccgg agaacaacta caagaccacg 960 cctcccgtgc tggactccga cggctccttc ttcctctaca gcaagctcac cgtggacaag 1020 agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggc tctgcacaac 1080 cactacacgc agaagagcct ctccctgtct ccgggtaaa 1119 <210> 20 <211> 373 <212> PRT <213> Homo sapiens <400> 20 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Phe Ser Asn Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Val Trp Val Ser Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr 65 70 75 80 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Ala Arg Trp Phe Gly Glu His Tyr Tyr Gly Met 115 120 125 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Glu Pro Lys 130 135 140 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 145 150 155 160 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 165 170 175 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 180 185 190 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 195 200 205 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 210 215 220 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 225 230 235 240 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 245 250 255 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 260 265 270 Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln 275 280 285 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 290 295 300 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 305 310 315 320 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 325 330 335 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 340 345 350 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 355 360 365 Leu Ser Pro Gly Lys 370 <210> 21 <211> 120 <212> PRT <213> Homo sapiens <400> 21 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45 Ser Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Phe Gly Glu His Tyr Tyr Gly Met Asp Val Trp Gly Gln<00013 / 06>100 105 110 Gly Thr Thr Val Thr Val Ser Ser / 15 120 <210> 22 <211> 10 <212> PRT <213> Homo sapiens <400> 22 Gly Phe Thr Phe Ser Asn Tyr Trp Met His 1 5 10 <210> 23 <211> 17 <212> PRT <213> Homo sapiens It should be noted that in the original text, "<00013 / 06>" seems to be an incorrect tag. It is likely a typo and should probably be " ". The above translation is based on the best understanding of the provided text. <400> 23 Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 24 <211> 11 <212> PRT <213> Homo sapiens <400> 24 Trp Phe Gly Glu His Tyr Tyr Gly Met Asp Val 1 5 10 <210> 25 <211> 1116 <212> DNA <213> Homo sapiens <400> 25 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga gggctgatcc agccgggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agaaacaact acatgagctg ggcccgccag 180 gttccaggga agggactgga gtgggtctca gttatttata gcggtgggag tataacctac 240 gcagactccg tgaagggccg attcatcatc tccagagaca attccaacaa cacgctatat 300 cttcaaatga acggtctgag agccgaggac acggccgtat attactgtgc gagagccgtc 360 ccacatgccc caagtgcttt tgacatctgg ggccagggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 26 <211> 372 <212> PRT <213> Homo sapiens <400> 26 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Arg Asn Asn Tyr Met Ser Trp Ala Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Asn 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 27 <211> 119 <212> PRT <213> Homo sapiens <400> 27 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Arg Asn Asn 20 25 30 Tyr Met Ser Trp Ala Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Asn Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 28 <211> 10 <212> PRT <213> Homo sapiens <400> 28 Gly Phe Thr Val Arg Asn Asn Tyr Met Ser 1 5 10 <210> 29 <211> 16 <212> PRT <213> Homo sapiens <400> 29 Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 30 <211> 11 <212> PRT <213> Homo sapiens <400> 30 Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile 1 5 10 <210> 31 <211> 1113 <212> DNA <213> Homo sapiens <400> 31 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga ggcttaatcc agcctgggg gtccctgaga 120 ctctcctgtg cagcctctgg gctcaatgtc agtaggcact acttaagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtcgca gtaatttata gcggtggtgg tagagttac 240 gtagactccg tgaagggccg attcaccatc tccagagaca atgccaacaa cacgctgtat 300 cttcaaatga gaagcctgag aaccgaagac tcggccgtat attactgtgc gcgggccatt 360 cccagacccc atggttcgga tatctggggc caagggacaa tggtcaccgt ctcttcagag 420 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 480 ggaccgtcag tcttctctctt ccccccaaaaa cccaaggaca ccctcatgat ctcccggacc 540 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 600 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 660 aacagcacgt accgggtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 720 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc 780 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc 780 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 acgcagaaga gcctctccct gtctccgggt aaa 1113 acgcagaaga gcctctccct gtctccgggt aaa 1113 <210> 32 <211> 371 <212> PRT <213> Homo sapiens <400> 32 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu 35 40 45 Asn Val Ser Arg His Tyr Leu Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ala Val Ile Tyr Ser Gly Gly Gly Ile Glu Tyr 65 70 75 80 Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Asn 85 90 95 Asn Thr Leu Tyr Leu Gln Met Arg Ser Leu Arg Thr Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Ile Pro Arg Pro His Gly Ser Asp Ile 115 120 125 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser Cys 130 135 140 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 145 150 155 160 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 165 170 175 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 180 185 190 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 195 200 205 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 210 215 220 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 225 230 235 240 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 245 250 255 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 260 265 270 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 275 280 285 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 290 295 300 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 305 310 315 320 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 325 330 335 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 340 345 350 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 355 360 365 Pro Gly Lys 370 <210> 33 <211> 118 <212> PRT <213> Homo sapiens <400> 33 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Asn Val Ser Arg His 20 25 30 Tyr Leu Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Tyr Ser Gly Gly Gly Ile Glu Tyr Val Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Asn Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Arg Ser Leu Arg Thr Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Ile Pro Arg Pro His Gly Ser Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 <210> 34 <211> 10 <212> PRT <213> Homo sapiens <400> 34 Gly Leu Asn Val Ser Arg His Tyr Leu Ser 1 5 10 <210> 35 <211> 16 <212> PRT <213> Homo sapiens <400> 35 Val Ile Tyr Ser Gly Gly Gly Ile Glu Tyr Val Asp Ser Val Lys Gly 1 5 10 15 <210> 36 <211> 10 <212> PRT <213> Homo sapiens <400> 36 Ala Ile Pro Arg Pro His Gly Ser Asp Ile 1 5 10 <210> 37 <211> 1116 <212> DNA <213> Homo sapiens <400> 37 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagcctctgg gttcaccgtc agtagcaact acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gttatttata gcggtggtag cacatactac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 300 cttcaaatga gcaacctgac cgccgaggac acggccgtgt attactgtgc gagagccgtt 360 ccccactccc ccagtgcttt tgataggtgg ggccagggaa ccctggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 38 <211> 372 <212> PRT <213> Homo sapiens <400> 38 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Ser Asn Leu Thr Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp 115 120 125 Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 39 <211> 119 <212> PRT <213> Homo sapiens <400> 39 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Ser Asn Leu Thr Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 40 <211> 10 <212> PRT <213> Homo sapiens <400> 40 Gly Phe Thr Val Ser Ser Asn Tyr Met Ser 1 5 10 <210> 41 <211> 16 <212> PRT <213> Homo sapiens <400> 41 Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 42 <211> 11 <212> PRT <213> Homo sapiens <400> 42 Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg 1 5 10 <210> 43 <211> 1116 <212> DNA <213> Homo sapiens <400> 43 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggagga gactggagga gacttgatcc agccgggggg gtccctgaga 120 ctctcctgta cagcctctgg gtttaacgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaggggccg atttaccatc tccagagaca gttccacgaa cacggtgtat 300 cttcaaatga acagcctgag agccgaggac tcggccgtgt attactgtgc gagagccgtc 360 ccacatgccc ctagtaattt tgataaatgg ggccaaggga caagggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 44<210> 44 <211> 372 <211> 372 <212> PRT <212> PRT <213> 智人 <213> Homo sapiens <400> 44 <400> 44 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe 35 40 45 Asn Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr 85 90 95 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp 115 120 125 Lys Trp Gly Gln Gly Thr Arg Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 45 <211> 119 <212> PRT <213> Homo sapiens <400> 45 Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Asn Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys Trp Gly Gln Gly 100 105 110 Thr Arg Val Thr Val Ser Ser 115 <210> 46 <211> 10 <212> PRT <213> Homo sapiens <400> 46 Gly Phe Asn Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 47 <211> 16 <212> PRT <213> Homo sapiens <400> 47 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 48 <211> 11 <212> PRT <213> Homo sapiens <400> 48 Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys 1 5 10 <210> 49 <211> 1125 <212> DNA <213> Homo sapiens <400> 49 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gtctggggga ggcttggttc agccgggggg gtccctgcga 120 ctctcctgtg tagcctctgg attcaccttc agtaactatt ggatgcactg ggtccgccaa 180 attccaggga aggggatggt gtggatctca gaaattaaca gtgacgggaa taggacaggc 240 tatgcggact ccgtaaaggg ccgattcacc atctccagag acaacgccaa gaagatgttg 300 tatctgcaaa tgaacagtct gagagccgag gacacggctg tttattattg tgcaagaaac 360 tggggatcgg gcccccatca taccgctttt gatggttggg gccaagggac aatggtcacc 420 gtctcttcag agcccaaatc ttgtgacaaa actcacacat gcccaccgtg cccagcacct 480 gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga caccctcatg 540 atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag 600 gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccgcgg 660 gaggagcagt acaacagcac gtaccgggtg gtcagcgtcc tcaccgtcct gcaccaggac 720 tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc 780 gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta caccctgccc 840 ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt caaaggcttc 900 tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag 960 accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg 1020 gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg 1080 cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaa 1125 <210> 50 <211> 375 <212> PRT <213> Homo sapiens <400> 50 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe 35 40 45 Thr Phe Ser Asn Tyr Trp Met His Trp Val Arg Gln Ile Pro Gly Lys 50 55 60 Gly Met Val Trp Ile Ser Glu Ile Asn Ser Asp Gly Asn Arg Thr Gly 65 70 75 80 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 85 90 95 Lys Lys Met Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 100 105 110 Ala Val Tyr Tyr Cys Ala Arg Asn Trp Gly Ser Gly Pro His His Thr 115 120 125 Ala Phe Asp Gly Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu 130 135 140 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 145 150 155 160 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 165 170 175 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 180 185 190 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 195 200 205 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 210 215 220 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 225 230 235 240 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 245 250 255 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 260 265 270 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 275 280 285 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 290 295 300 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 305 310 315 320 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser​​​​​​​​​​​​​​​​​​​​​​​​​Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ile Pro Gly Lys Gly Met Val Trp Ile 35 40 45 Ser Glu Ile Asn Ser Asp Gly Asn Arg Thr Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Met Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Trp Gly Ser Gly Pro His His Thr Ala Phe Asp Gly Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 52 <211> 10 <212> PRT <213> Homo sapiens <400> 52 Gly Phe Thr Phe Ser Asn Tyr Trp Met His 1 5 10 <210> 53 <211> 17 <212> PRT <213> Homo sapiens <400> 53 Glu Ile Asn Ser Asp Gly Asn Arg Thr Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 54 <211> 13 <212> PRT <213> Homo sapiens <400> 54 Asn Trp Gly Ser Gly Pro His His Thr Ala Phe Asp Gly 1 5 10 <210> 55 <211> 1116 <212> DNA <213> Homo sapiens <400> 55 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggagga gactggagga gacttgatcc agccgggggg gtccctgaga 120 ctctcctgta cagcctctgg gtttaccgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaggggccg atttaccatc tccagagaca gttccacgaa cacggtgttt 300 cttcaaatga acagcctgag agccgaggac tcggccgtgt attactgtgc gagagccgtc 360 ccaagtgccc ctagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 56 <211> 372 <212> PRT <213> Homo sapiens <400> 56 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe 35 40 45 Thr Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr 85 90 95 Asn Thr Val Phe Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 57 <211> 119 <212> PRT <213> Homo sapiens <400> 57 Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr Asn Thr Val Phe Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 58 <211> 10 <212> PRT <213> Homo sapiens <400> 58 Gly Phe Thr Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 59 <211> 16 <212> PRT <213> Homo sapiens <400> 59 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 60 <211> 11 <212> PRT <213> Homo sapiens <400> 60 Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 1 5 10 <210> 61 <211> 1116 <212> DNA <213> Homo sapiens <400> 61 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gagcggagga ggcttgatcc agccgggggg gtccctgaga 120 ctctcctgtg ccgcctctgg gtttaccgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaagggccg atttaccatc tccagagaca gttccaagaa cacggtgtac 300 cttcaaatga acagcctgag agccgaggac accgccgtgt attactgtgc gagagccgtc 360 ccaagtgccc ctagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 62 <211> 372 <212> PRT <213> Homo sapiens <400> 62 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys 85 90 95 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 63 <211> 119 <212> PRT <213> Homo sapiens <400> 63 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 64 <211> 10 <212> PRT <213> Homo sapiens <400> 64 Gly Phe Thr Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 65 <211> 16 <212> PRT <213> Homo sapiens <400> 65 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 66 <211> 11 <212> PRT <213> Homo sapiens <400> 66 Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 1 5 10 <210> 67 <211> 1116 <212> DNA <213> Homo sapiens <400> 67 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga gggctgatcc agccgggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agaaacaact acatgagctg ggcccgccag 180 gttccaggga agggactgga gtgggtctca gttatttata gcggtgggag tataacctac 240 gcagactccg tgaagggccg attcatcatc tccagagaca attccaagaa cacgctatat 300 cttcaaatga acggtctgag agccgaggac acggccgtat attactgtgc gagagccgtc 360 ccacatgccc caagtgcttt tgacatctgg ggccagggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 68 <211> 372 <212> PRT <213> Homo sapiens <400> 68 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Arg Asn Asn Tyr Met Ser Trp Ala Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 69 <211> 119 <212> PRT <213> Homo sapiens <400> 69 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Arg Asn Asn 20 25 30 Tyr Met Ser Trp Ala Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 70 <211> 10 <212> PRT <213> Homo sapiens <400> 70 Gly Phe Thr Val Arg Asn Asn Tyr Met Ser 1 5 10 <210> 71 <211> 16 <212> PRT <213> Homo sapiens <400> 71 Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 72 <211> 11 <212> PRT <213> Homo sapiens <400> 72 Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile 1 5 10 <210> 73 <211> 1116 <212> DNA <213> Homo sapiens <400> 73 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gagcggagga gggctgatcc agccgggggg gtccctgaga 120 ctctcctgtg cagcctctgg gttcaccgtc agaaacaact acatgagctg ggtgcgccag 180 gccccaggga agggactgga gtgggtctca gttatttata gcggtgggag tataacctac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctatat 300 cttcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gagagccgtc 360 ccacatgccc caagtgcttt tgacatctgg ggccagggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 74 <211> 372 <212> PRT <213> Homo sapiens <400> 74 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Arg Asn Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 75 <211> 119 <212> PRT <213> Homo sapiens <400> 75 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Arg Asn Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 76 <211> 10 <212> PRT <213> Homo sapiens[[ID=ip=19]] <400> 76 Gly Phe Thr Val Arg Asn Asn Tyr Met Ser 1 5 10 <210> 77 <211> 16 <212> PRT <213> Homo sapiens <400> 77 Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 78 [[ID=4ip==2]]<211> 11 <212> PRT <213> Homo sapiens <400> 78 Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile 1 5 10 <210> 79 <211> 1116 <212> DNA <213> Homo sapiens <400> 79 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga ggctgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagcctctgg gttcaccgtc agtagcact acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gttatta gcggtggtag cacatactac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 300 cttcaaatga gcagcctgag ggccgaggac acggccgtgt attactgtgc gagagccgtt 360 ccccactccc ccagtgcttt tgatagtgg ggccagga ccctgtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccaccacc tgaactcctg 480 gggggaccgt cagtctctcct cttcccccca aaacccaagg acacctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg agaccctga ggtcaagttc 600 aactggtacg tggacgggt ggaggtgcat aatgccaaga caagccgcg ggaggagcag 660 tacacagca cgtaccggggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaggagt acagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 80 <211> 372 <212> PRT <213> Homo sapiens <400> 80 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp 115 120 125 Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 81 <211> 119 <212> PRT <213> Homo sapiens <400> 81 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 82 <211> 10 <212> PRT <213> Homo sapiens <400> 82 Gly Phe Thr Val Ser Ser Asn Tyr Met Ser 1 5 10 <210> 83 <211> 16 <212> PRT <213> Homo sapiens <400> 83 Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 84 <211> 11 <212> PRT <213> Homo sapiens <400> 84 Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg 1 5 10 <210> 85 <211> 1116 <212> DNA <213> Homo sapiens <400> 85 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gagcggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagcctctgg gttcaccgtc agtagcaact acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gttatttata gcggtggtag cacatactac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 300 cttcaaatga acagcctgag ggccgaggac acggccgtgt attactgtgc gagagccgtt 360 ccccactccc ccagtgcttt tgataggtgg ggccagggaa ccctggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 86 <211> 372 <212> PRT <213> Homo sapiens <400> 86 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp 115 120 125 Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 87 <211> 119 <212> PRT <213> Homo sapiens <400> 87 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 88 <211> 10 <212> PRT <213> Homo sapiens <400> 88 Gly Phe Thr Val Ser Ser Asn Tyr Met Ser 1 5 10 <210> 89 <211> 16 <212> PRT <213> Homo sapiens <400> 89 Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 90 <211> 11 <212> PRT <213> Homo sapiens <400> 90 Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg 1 5 10 <210> 91 <211> 1116 <212> DNA <213> Homo sapiens <400> 91 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggagga gactggagga gacttgatcc agccgggggg gtccctgaga 120 ctctcctgta cagcctctgg gtttaccgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaggggccg atttaccatc tccagagaca gttccacgaa cacggtgtat 300 cttcaaatga acagcctgag agccgaggac tcggccgtgt attactgtgc gagagccgtc 360 ccacatgccc ctagtaattt tgataaatgg ggccaaggga caagggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 92 <211> 372 <212> PRT <213> Homo sapiens <400> 92 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe 35 40 45 Thr Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr 85 90 95 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp 115 120 125 Lys Trp Gly Gln Gly Thr Arg Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 93 <211> 119 <212> PRT <213> Homo sapiens <400> 93 Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys Trp Gly Gln Gly 100 105 110 Thr Arg Val Thr Val Ser Ser 115 <210> 94 <211> 10 <212> PRT <213> Homo sapiens <400> 94 Gly Phe Thr Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 95 <211> 16 <212> PRT <213> Homo sapiens <400> 95 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 96 <211> 11 <212> PRT <213> Homo sapiens <400> 96 Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys 1 5 10 <210> 97 <211> 1116 <212> DNA <213> Homo sapiens <400> 97 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggcgga gactggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agtaagaatt acatgagctg ggtccgccag 180 gttccaggga aggggctgga gtgggtcgca gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcatcctc tccagagaca attccgacaa cacactgtat 300 cttcaaatga acaacctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 98 <211> 372 <212> PRT <213> Homo sapiens <400> 98 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys 85 90 95 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp 115 120 125 Lys Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 99 <211> 119 <212> PRT <213> Homo sapiens <400> 99 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 100 <211> 10 <212> PRT <213> Homo sapiens <400> 100 Gly Phe Thr Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 101 <211> 16 <212> PRT <213> Homo sapiens <400> 101 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 102 <211> 11 <212> PRT <213> Homo sapiens <400> 102 Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys 1 5 10 <210> 103 <211> 1116 <212> DNA <213> Homo sapiens <400> 103 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggcgga gactggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agtaagaatt acatgagctg ggtccgccag 180 gttccaggga aggggctgga gtgggtcgca gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcatcctc tccagagaca attccgacaa cacactgtat 300 cttcaaatga acaacctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 104 <211> 372 <212> PRT <213> Homo sapiens <400> 104 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 105 <211> 119 <212> PRT <213> Homo sapiens <400> 105 Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 106 <211> 10 <212> PRT <213> Homo sapiens <400> 106 Gly Phe Thr Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 107 <211> 16 <212> PRT <213> Homo sapiens <400> 107 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 108 <211> 11 <212> PRT <213> Homo sapiens <400> 108 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 109 <211> 1116 <212> DNA <213> Homo sapiens <400> 109 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gagcggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agtaagaatt acatgagctg ggtccgccag 180 gccccaggga aggggctgga gtgggtcagc gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtat 300 cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 110 <211> 372 <212> PRT <213> Homo sapiens <400> 110 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 111 <211> 119 <212> PRT <213> Homo sapiens <400> 111 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 112 <211> 10 <212> PRT <213> Homo sapiens <400> 112 Gly Phe Thr Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 113 <211> 16 <212> PRT <213> Homo sapiens <400> 113 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 114 <211> 11 <212> PRT <213> Homo sapiens <400> 114 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 115 <211> 1116 <212> DNA <213> Homo sapiens <400> 115 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggagga gactggagga gacttgatcc agccgggggg gtccctgaga 120 ctctcctgta cagcctctgg gtttaacgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaggggccg atttaccatc tccagagaca gttccacgaa cacggtgttt 300 cttcaaatga acagcctgag agccgaggac tcggccgtgt attactgtgc gagagccgtc 360 ccaagtgccc ctagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 116 <211> 372 <212> PRT <213> Homo sapiens <400> 116 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe 35 40 45 Asn Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr 85 90 95 Asn Thr Val Phe Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 117 <211> 119 <212> PRT <213> Homo sapiens <400> 117 Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Asn Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr Asn Thr Val Phe Leu 65 70 7 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 118 <211> 10 <212> PRT <213> Homo sapiens <400> 118 Gly Phe Asn Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 119 <211> 16 <212> PRT <213> Homo sapiens <400> 119 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 120 <211> 11 <212> PRT <213> Homo sapiens <400> 120 Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 1 5 10 <210> 121 <211> 1116 <212> DNA <213> Homo sapiens <400> 121 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga gggctgatcc agccgggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agaaacaact acatgagctg ggcccgccag 180 gttccaggga agggactgga gtgggtctca gttatttata gcggtgggag tataacctac 240 gcagactccg tgaagggccg attcatcatc tccagagaca attccaacaa cacgctatat 300 cttcaaatga acggtctgag agccgaggac acggccgtat attactgtgc gagagccgtc 360 ccacatgccc caagtgcttt tgacatctgg ggccagggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 122 <211> 372 <212> PRT <213> Homo sapiens <400> 122 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Arg Asn Asn Tyr Met Ser Trp Ala Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Asn 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 123 <211> 119 <212> PRT <213> Homo sapiens <400> 123 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Arg Asn Asn 20 25 30 Tyr Met Ser Trp Ala Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Asn Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 124 <211> 10 <212> PRT <213> Homo sapiens <400> 124 Gly Phe Thr Val Arg Asn Asn Tyr Met Ser 1 5 10 <210> 125 <211> 16 <212> PRT <213> Homo sapiens <400> 125 Val Ile Tyr Ser Gly Gly Ser Ile Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 126 <211> 11 <212> PRT <213> Homo sapiens <400> 126 Ala Val Pro His Ala Pro Ser Ala Phe Asp Ile 1 5 10 <210> 127 <211> 1116 <212> DNA <213> Homo sapiens <400> 127 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gactggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagcctctgg gttcaccgtc agtagcaact acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gttatta gcggtggtag cacatactac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 300 cttcaaatga gcaacctgac cgccgaggac acggccgtgt attackgtgc gagagccgtt 360 ccccactccc ccagtgcttt tgatagtgg ggccagga ccctgtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccaccacc tgaactcctg 480 gggggaccgg acgtctctcct cttcccccca aaacccaagg acacctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg agaccctga ggtcaagttc 600 aactggtacg tggacgggt ggaggtgcat aatgccaaga caagccgcg ggaggagcag 660 tacacagca cgtaccggggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaggagt acagtgcaa ggtctccac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca gtcagcctg acctgcctgg tcaaggctt ctacccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 128 <211> 372 <212> PRT <213> Homo sapiens <400> 128 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 35 40 45 Thr Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Ser Asn Leu Thr Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp 115 120 125 Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 129 <211> 119 <212> PRT <213> Homo sapiens <400> 129 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Ser Asn Leu Thr Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 130 <211> 10 <212> PRT <213> Homo sapiens <400> 130 Gly Phe Thr Val Ser Ser Asn Tyr Met Ser 1 5 10 <210> 131 <211> 16 <212> PRT <213> Homo sapiens <400> 131 Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 132 <211> 11 <212> PRT <213> Homo sapiens <400> 132 Ala Val Pro His Ser Pro Ser Ala Phe Asp Arg 1 5 10 <210> 133 <211> 1116 <212> DNA <213> Homo sapiens <400> 133 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggagga gactggagga gacttgatcc agccgggggg gtccctgaga 120 ctctcctgta cagcctctgg gtttaacgtc agtagaatct acatgagctg ggtccgccag 180 gctccaggga aggggctgga gtgggtctca gtgatttata gcggtggaag tatatattac 240 gcagattccg tgaggggccg atttaccatc tccagagaca gttccacgaa cacggtgtat 300 cttcaaatga acagcctgag agccgaggac tcggccgtgt attactgtgc gagagccgtc 360 ccacatgccc ctagtaattt tgataaatgg ggccaaggga caagggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 134 <211> 372 <212> PRT <213> Homo sapiens <400> 134 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe 35 40 45 Asn Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr 65 70 75 80 Ala Asp Ser Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr 85 90 95 Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp 115 120 125 Lys Trp Gly Gln Gly Thr Arg Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 135 <211> 119 <212> PRT <213> Homo sapiens <400> 135 Glu Val Gln Leu Glu Glu Thr Gly Gly Asp Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Asn Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Thr Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys Trp Gly Gln Gly 100 105 110 Thr Arg Val Thr Val Ser Ser 115 <210> 136 <211> 10 <212> PRT <213> Homo sapiens <400> 136 Gly Phe Asn Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 137 <211> 16 <212> PRT <213> Homo sapiens <400> 137 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 138 <211> 11 <212> PRT <213> Homo sapiens <400> 138 Ala Val Pro His Ala Pro Ser Asn Phe Asp Lys 1 5 10 <210> 139 <211> 1116 <212> DNA <213> Homo sapiens <400> 139 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggcgga gactggagga ggcttgatcc agcctgggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaatgtc agtaagaatt acatgagctg ggtccgccag 180 gttccaggga aggggctgga gtgggtcgca gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcatcctc tccagagaca attccgacaa cacactgtat 300 cttcaaatga aaacctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 140 <211> 372 <212> PRT <213> Homo sapiens <400> 140 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Asn Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 141 <211> 119 <212> PRT <213> Homo sapiens <400> 141 Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Asn Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 142 <211> 10 <212> PRT <213> Homo sapiens <400> 142 Gly Phe Asn Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 143 <211> 16 <212> PRT <213> Homo sapiens <400> 143 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 144 <211> 11 <212> PRT <213> Homo sapiens <400> 144 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 145 <211> 1116 <212> DNA <213> Homo sapiens <400> 145 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggcgga gactggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agtaagaatt acatgagctg ggtccgccag 180 gttccaggga aggggctgga gtgggtcgca gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcatcctc tccagagaca attccgacaa cacactgtat 300 cttcaaatga acaacctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 146 <211> 372 <212> PRT <213> Homo sapiens <400> 146 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 147 <211> 119 <212> PRT <213> Homo sapiens <400> 147 Glu Val Gln Leu Ala Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ile Leu Ser Arg Asp Asn Ser Asp Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 148 <211> 10 <212> PRT <213> Homo sapiens <400> 148 Gly Phe Thr Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 149 <211> 16 <212> PRT <213> Homo sapiens <400> 149 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 150 <211> 11 <212> PRT <213> Homo sapiens <400> 150 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 151 <211> 1116 <212> DNA <213> Homo sapiens <400> 151 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 tctgaggtgc agctggtgga gagcggagga ggcttgatcc agcctggggg gtccctgaga 120 ctctcctgtg cagtctctgg gttcaccgtc agtaagaatt acatgagctg ggtccgccag 180 gccccaggga aggggctgga gtgggtcagc gttgtttata gcggtggtag caaaacctac 240 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtat 300 cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagagccgtt 360 cctcattccc ccagttcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 152 <211> 372 <212> PRT <213> Homo sapiens <400> 152 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 20 25 30 Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe 35 40 45 Thr Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys 50 55 60 Gly Leu Glu Trp Val Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp 115 120 125 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 153 <211> 119 <212> PRT <213> Homo sapiens <400> 153 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 154 <211> 10 <212> PRT <213> Homo sapiens <400> 154 Gly Phe Thr Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 155 <211> 16 <212> PRT <213> Homo sapiens <400> 155 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 156 <211> 11 <212> PRT <213> Homo sapiens <400> 156 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 157 <211> 1116 <212> DNA <213> Homo sapiens <400> 157 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 gaggtgcagc tggtggagtc cgggggaggc ttagttcagc ctggggggtc cctgagactc 120 tcctgtgcag cctctggatt caccttcagt aactactgga tgcactgggt ccgccaagct 180 ccagggaagg ggctggtgtg ggtctcacgt attaatagtg atgggagtaa cacaacctac 240 gcggactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacgctgtat 300 ctgcaaatga acagtctgag agccgaggac acggctgtgt attactgtgc aagatggttc 360 ggggagcact actacggaat ggacgtctgg ggccaaggga ccacggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 158 <211> 372 <212> PRT <213> Homo sapiens <400> 158 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Asn Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Val Trp Val Ser Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Trp Phe Gly Glu His Tyr Tyr Gly Met Asp 115 120 125 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 159 <211> 120 <212> PRT <213> Homo sapiens <400> 159 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45 Ser Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Phe Gly Glu His Tyr Tyr Gly Met Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 160 <211> 10 <212> PRT <213> Homo sapiens <400> 160 Gly Phe Thr Phe Ser Asn Tyr Trp Met His 1 5 10 <210> 161 <211> 17 <212> PRT <213> Homo sapiens <400> 161 Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 162 <211> 11 <212> PRT <213> Homo sapiens <400> 162 Trp Phe Gly Glu His Tyr Tyr Gly Met Asp Val 1 5 10 <210> 163 <211> 1113 <212> DNA <213> Homo sapiens <400> 163 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 gaggtgcagc tggtggagag cggaggaggc ttgatccagc cgggggggtc cctgagactc 120 tcctgtgccg cctctgggtt taccgtcagt agaatctaca tgagctgggt ccgccaggct 180 ccagggaagg ggctggagtg ggtctcagtg atttatagcg gtggaagtat atattacgca 240 gattccgtga agggccgatt taccatctcc agagacagtt ccaagaacac ggtgtacctt 300 caaatgaaca gcctgagagc cgaggacacc gccgtgtatt actgtgcgag agccgtccca 360 agtgccccta gttcttttga tatctggggc caagggacaa tggtcaccgt ctcttcagag 420 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 480 ggaccgtcag tcttctctctt ccccccaaaaa cccaaggaca ccctcatgat ctcccggacc 540 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 600 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 660 aacagcacgt accgggtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 720 aaggagtca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaccatc 780 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 your acgcaga gcctctccct gtctccgggt aaa 1113 <210> 164 <211> 371 <212> PRT <213> Homo sapiens <400> 164 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn 85 90 95 Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 115 120 125 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser Cys 130 135 140 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 145 150 155 160 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 165 170 175 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 180 185 190 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 195 200 205 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 210 215 220 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 225 230 235 240 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 245 250 255 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 260 265 270 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 275 280 285 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 290 295 300 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 305 310 315 320 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 325 330 335 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 340 345 350 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 355 360 365 Pro Gly Lys 370 <210> 165 <211> 119 <212> PRT <213> Homo sapiens <400> 165 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 166 <211> 10 <212> PRT <213> Homo sapiens <400> 166 Gly Phe Thr Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 167 <211> 16 <212> PRT <213> Homo sapiens <400> 167 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 168 <211> 11 <212> PRT <213> Homo sapiens <400> 168 Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 1 5 10 <210> 169 <211> 1113 <212> DNA <213> Homo sapiens <400> 169 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 gaggtgcagc tggtggagag cggaggaggc ttgatccagc ctggggggtc cctgagactc 120 tcctgtgcag tctctgggtt caccgtcagt aagaattaca tgagctgggt ccgccaggcc 180 ccagggaagg ggctggagagtg ggtcagcgtt gtttatagcg gtggtagcaa aacctacgca 240 gactccgtga agggccgatt caccatctcc agagacaatt ccaagaacac actgtatctt 300 caaatgaaca gcctgagagc cgaggacacg gccgtgtatt actgtgcgag agccgttcct 360 cattccccca gttcttttga tatctggggc caagggacaa tggtcaccgt ctcttcagag 420 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 480 ggaccgtcag tcttctctctt ccccccaaaaa cccaaggaca ccctcatgat ctcccggacc 540 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 600 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 660 aacagcacgt accgggtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 720 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc 780 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 acgcagaaga gcctctccct gtctccgggt aaa 1113 <210> 170 <211> 371 <212> PRT <213> Homo sapiens <400> 170 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr 35 40 45 Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 85 90 95 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 115 120 125 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser Cys 130 135 140 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 145 150 155 160 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 165 170 175 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 180 185 190 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 195 200 205 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 210 215 220 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 225 230 235 240 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 245 250 255 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 260 265 270 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 275 280 285 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 290 295 300 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 305 310 315 320 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 325 330 335 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 340 345 350 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 355 360 365 Pro Gly Lys 370 <210> 171 <211> 119 <212> PRT <213> Homo sapiens <400> 171 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 172 <211> 10 <212> PRT <213> Homo sapiens <400> 172 Gly Phe Thr Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 173 <211> 16 <212> PRT <213> Homo sapiens <400> 173 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 174 <211> 11 <212> PRT <213> Homo sapiens <400> 174 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 175 <211> 1113 <212> DNA <213> Homo sapiens <400> 175 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 gaggtgcagc tggtggagag cggaggaggc ttgatccagc cgggggggtc cctgagactc 120 tcctgtgccg cctctgggtt taccgtcagt agaatctaca tgagctgggt ccgccaggct 180 ccagggaagg ggctggagtg ggtctcagtg atttatagcg gtggaagtat atattacgca 240 gattccgtga agggccgatt taccatctcc agagacagtt ccaagaacac ggtgtacctt 300 aaatgaaca gcctgagagc cgaggacacc gccgtgtatt actgtgcgag agccgtccca 360 420 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 480 ggaccggacg tcttctctctt ccccccaaaaa cccaaggaca ccctcatgat ctcccggacc 540 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 600 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 660 aacagcacgt accgggtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 720 aaggagtca agtgcaaggt ctccaacaaa gccctcccag cccccgagga gaaaccatc 780 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 acgcagaaga gcctctccct gtctccgggt aaa 1113 <210> 176 <211> 371 <212> PRT <213> Homo sapiens <400> 176 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Val Ser Arg Ile Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn 85 90 95 Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 115 120 125 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser Cys 130 135 140 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 145 150 155 160 Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 165 170 175 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 180 185 190 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 195 200 205 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 210 215 220 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 225 230 235 240 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu 245 250 255 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 260 265 270 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 275 280 285 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 290 295 300 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 305 310 315 320 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 325 330 335 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 340 345 350 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 355 360 365 Pro Gly Lys 370 <210> 177 <211> 119 <212> PRT <213> Homo sapiens <400> 177 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Arg Ile 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 178 <211> 10 <212> PRT <213> Homo sapiens <400> 178 Gly Phe Thr Val Ser Arg Ile Tyr Met Ser 1 5 10 <210> 179 <211> 16 <212> PRT <213> Homo sapiens <400> 179 Val Ile Tyr Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 180 <211> 11 <212> PRT <213> Homo sapiens <400> 180 Ala Val Pro Ser Ala Pro Ser Ser Phe Asp Ile 1 5 10 <210> 181 <211> 1113 <212> DNA <213> Homo sapiens <400> 181 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 gaggtgcagc tggtggagag cggaggaggc ttgatccagc ctggggggtc cctgagactc 120 tcctgtgcag tctctgggtt caccgtcagt aagaattaca tgagctgggt ccgccaggcc 180 ccagggaagg ggctggagtg ggtcagcgtt gtttatagcg gtggtagcaa aacctacgca 240 gactccgtga agggccgatt caccatctcc agagacaatt ccaagaacac actgtatctt 300 caaatgaaca gcctgagagc cgaggacacg gccgtgtatt actgtgcgag agccgttcct 360 cattccccca gttcttttga tatctggggc caagggacaa tggtcaccgt ctcttcagag 420 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg 480 ggaccggacg tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 540 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 600 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 660 aacagcacgt accgggtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 720 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccgagga gaaaaccatc 780 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 840 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 900 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 960 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 1020 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 1080 acgcagaaga gcctctccct gtctccgggt aaa 1113 <210> 182 <211> 371 <212> PRT <213> Homo sapiens <400> 182 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr 35 40 45 Val Ser Lys Asn Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 85 90 95 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 115 120 125 Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Glu Pro Lys Ser Cys 130 135 140 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 145 150 155 160 Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 165 170 175 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 180 185 190 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 195 200 205 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 210 215 220 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 225 230 235 240 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu 245 250 255 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 260 265 270 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 275 280 285 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 290 295 300 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 305 310 315 320 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 325 330 335 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 340 345 350 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 355 360 365 Pro Gly Lys 370 <210> 183 <211> 119 <212> PRT <213> Homo sapiens <400> 183 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Ser Lys Asn 20 25 30 Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 184 <211> 10 <212> PRT <213> Homo sapiens <400> 184 Gly Phe Thr Val Ser Lys Asn Tyr Met Ser 1 5 10 <210> 185 <211> 16 <212> PRT <213> Homo sapiens <400> 185 Val Val Tyr Ser Gly Gly Ser Lys Thr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 186 <211> 11 <212> PRT <213> Homo sapiens <400> 186 Ala Val Pro His Ser Pro Ser Ser Phe Asp Ile 1 5 10 <210> 187 <211> 1116 <212> DNA <213> Homo sapiens <400> 187 atggagaccg acaccctgct gctgtgggtg ctgctgctgt gggtgcccgg atccaccggc 60 gaggtgcagc tggtggagtc cgggggaggc ttagttcagc ctggggggtc cctgagactc 120 tcctgtgcag cctctggatt caccttcagt aactactgga tgcactgggt ccgccaagct 180 ccagggaagg ggctggtgtg ggtctcacgt attaatagtg atgggagtaa cacaacctac 240 gcggactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacgctgtat 300 ctgcaaatga acagtctgag agccgaggac acggctgtgt attactgtgc aagatggttc 360 ggggagcact actacggaat ggacgtctgg ggccaaggga ccacggtcac cgtctcttca 420 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 480 gggggaccgg acgtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 540 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 600 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 660 tacaacagca cgtaccgggt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 720 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccga ggagaaaacc 780 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 840 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 900 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 960 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1020 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1080 tacacgcaga agagcctctc cctgtctccg ggtaaa 1116 <210> 188 <211> 372 <212> PRT <213> Homo sapiens <400> 188 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Asn Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Val Trp Val Ser Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Trp Phe Gly Glu His Tyr Tyr Gly Met Asp 115 120 125 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Glu Pro Lys Ser 130 135 140 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 145 150 155 160 Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 165 170 175 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 180 185 190 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 195 200 205 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 210 215 220 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 225 230 235 240 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 245 250 255 Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 260 265 270 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 275 280 285 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 290 295 300 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 305 310 315 320 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 325 330 335 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 340 345 350 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 355 360 365 Ser Pro Gly Lys 370 <210> 189 <211> 120 <212> PRT <213> Homo sapiens <400> 189 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45 Ser Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Phe Gly Glu His Tyr Tyr Gly Met Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 190 <211> 10 <212> PRT <213> Homo sapiens <400> 190 Gly Phe Thr Phe Ser Asn Tyr Trp Met His 1 5 10 <210> 191 <211> 17 <212> PRT <213> Homo sapiens <400> 191 Arg Ile Asn Ser Asp Gly Ser Asn Thr Thr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 192 <211> 11 <212> PRT <213> Homo sapiens <400> 192 Trp Phe Gly Glu His Tyr Tyr Gly Met Asp Val 1 5 10 <210> 193 <211> twenty three <212> DNA <213> Artificial sequence <220> <223> Primers <400> 193 gtgtccagtg tgaggtgcag ctg 23 <210> 194 <211> twenty three <212> DNA <213> Artificial sequence <220> <223> Primers <400> 194 gtgtccagtg tcaggtgcag ctg 23 <210> 195 <211> twenty four <212> DNA <213> Artificial sequence <220> <223> Primers <400> 195 ggcttacaac cacaatccct gggc 24 <210> 196 <211> 672 <212> DNA <213> Homo sapiens <400> 196 atggcttgcc ttggatttca gcggcacaag gctcagctga acctggctac caggacctgg 60 ccctgcactc tcctgttttt tcttctcttc atccctgtct tctgcaaagc aatgcacgtg 120 gcccagcctg ctgtggtact ggccagcagc cgaggcatcg ccagctttgt gtgtgagtat 180 gcatctccag gcaaagccac tgaggtccgg gtgacagtgc ttcggcaggc tgacagccag 240 gtgactgaag tctgtgcggc aacctacatg atggggaatg agttgacctt cctagatgat 300 tccatctgca cgggcacctc cagtggaaat caagtgaacc tcactatcca aggactgagg 360 gccatggaca cgggactcta catctgcaag gtggagctca tgtacccacc gccatactac 420 ctgggcatag gcaacggaac ccagatttat gtaattgatc cagaaccgtg cccagattct 480 gacttcctcc tctggatcct tgcagcagtt agttcggggt tgttttttta tagctttctc 540 ctcacagctg tttctttgag caaaatgcta aagaaaagaa gccctcttac aacaggggtc 600 tatgtgaaaa tgcccccaac agagccagaa tgtgaaaagc aatttcagcc ttattttatt 660 cccatca...

Claims

1. An isolated monoclonal antibody comprising a CD152 binding domain, wherein the CD152 binding domain comprises an immunoglobulin heavy chain variable region comprising CDR1, CDR2 and CDR3, wherein the amino acid sequences of CDR1, CDR2 and CDR3 are respectively as shown below: (1) SEQ ID NO: 16, 17 and 18; (2) SEQ ID NO: 58, 59 and 60; or (3) SEQ ID NO: 64, 65 and 66, wherein the antibody does not comprise an immunoglobulin light chain.

2. The isolated monoclonal antibody of claim 1, wherein the antibody is a heavy chain only antibody.

3. The isolated monoclonal antibody of claim 1 or 2, wherein the antibody comprises two immunoglobulin heavy chains.

4. The isolated monoclonal antibody of any one of claims 1 to 3, wherein the antibody consists of two immunoglobulin heavy chains.

5. The isolated monoclonal antibody of any one of claims 1 to 4, wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 15, 57 or 63.

6. The isolated monoclonal antibody of claim 5, wherein the immunoglobulin heavy chain variable domain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 15, 57 or 63.

7. The isolated monoclonal antibody of claim 5, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence shown in SEQ ID NO: 15, 57 or 63.

8. The isolated monoclonal antibody of any one of claims 3 to 7, wherein at least one of the two immunoglobulin heavy chains comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 14, 56, 62, 116, 164 or 176.

9. The isolated monoclonal antibody of claim 8, wherein at least one of the two immunoglobulin heavy chains comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 14, 56, 62, 116, 164 or 176.

10. The isolated monoclonal antibody of claim 8, wherein at least one of the two immunoglobulin heavy chains comprises the amino acid sequence shown in SEQ ID NO: 14, 56, 62, 116, 164 or 176.

11. The isolated monoclonal antibody of any one of claims 1 to 10, which exhibits one or a combination of the following properties: (a) binding to human CD152; (b) specific binding to monkey CD152; (c) not binding to mouse CD152; (d) blocking the binding of CD152 to CD80, CD86 or both; (e) promoting IL-2 secretion by immune cells; (f) inducing T cell activation; (g) stimulating anti-tumor immune response of immune cells.

12. The isolated monoclonal antibody of any one of claims 1 to 11, which exhibits one or a combination of the following properties: (a) binds to human CD152 with an affinity higher than that of an ipilimumab analog; (b) binds to human CD152 with an affinity of 1.0*10 -9 M or smaller K d Dissociation from human CD152; (c) K compared to ipilimumab analogs d Lower K d Dissociated from human CD152, wherein the ipilimumab analog comprises a heavy chain having an amino acid sequence of SEQ ID NO: 199 and a light chain having an amino acid sequence of SEQ ID NO:

200.

13. The isolated monoclonal antibody of any one of claims 1 to 12, wherein the antibody is expressed at 6.0*10 -11 M or smaller K d Dissociated from human CD152.

14. The isolated monoclonal antibody of any one of claims 1 to 13, wherein the antibody is a human antibody, a humanized antibody, or a chimeric antibody.

15. A pharmaceutical composition comprising the isolated monoclonal antibody of any one of claims 1 to 14 and a pharmaceutically acceptable excipient.

16. The pharmaceutical composition of claim 15, wherein the pharmaceutically acceptable excipient is selected from the group consisting of a carrier, a surfactant, a thickener or emulsifier, a solid binder, a dispersion or suspension aid, a solubilizer, a colorant, a flavoring agent, a coating, a disintegrant, a lubricant, a sweetener, a preservative, an isotonic agent, and a combination thereof.

17. The pharmaceutical composition of claim 15 or 16, further comprising a second antibody, wherein the second antibody is an immunostimulatory antibody or a co-stimulatory antibody.

18. A nucleic acid molecule encoding the isolated monoclonal antibody of any one of claims 1 to 14.

19. The nucleic acid molecule of claim 18, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 90% identity to SEQ ID NO: 13, 55, 61, 115, 163 or 175.

20. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 95% identity to SEQ ID NO: 13, 55, 61, 115, 163 or 175.

21. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO: 13, 55, 61, 115, 163 or 175.

22. An expression vector comprising the nucleic acid molecule of any one of claims 18 to 21, wherein the nucleic acid molecule is operably linked to regulatory sequences suitable for expressing the nucleic acid segment in a host cell.

23. A host cell comprising the expression vector of claim 22.

24. Use of the isolated monoclonal antibody of any one of claims 1 to 14 or the pharmaceutical composition of any one of claims 15 to 17 in the preparation of a medicament for treating a disorder, wherein the disorder is a solid tumor.

25. The method of claim 24, wherein the solid tumor is selected from the group consisting of renal cell carcinoma, colorectal cancer, breast cancer, epithelial squamous cell carcinoma, melanoma, myeloma, gastric cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, and head and neck cancer.

26. The use of claim 24, wherein the solid tumor is colon cancer.

27. The use according to any one of claims 24 to 26, wherein the medicament further comprises an additional therapeutic agent.

28. The use of claim 27, wherein the additional therapeutic agent is an anti-cancer agent.

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