Monoclonal antibodies against her2 epitope

Abstract

Isolated monoclonal antibodies which bind to human epidermal growth factor receptor 2 (HER2), and related antibody-based compositions and molecules, are disclosed. Pharmaceutical compositions comprising the antibodies and therapeutic and diagnostic methods for using the antibodies are also disclosed.

1 - 55 . (canceled) 
     
     
         56 . A method for inhibiting the growth and/or proliferation of one or more tumor cells expressing human epidermal growth factor receptor 2 (HER2) comprising administering to an individual in need thereof an effective amount of an antibody comprising a variable heavy chain (VH) region and a variable light chain (VL) region selected from the group consisting of:
 (a) an antibody comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 2, 3 and 4, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 6, GAS, and SEQ ID NO: 7, respectively;   (b) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 9, 10 and 11, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 13, DAS, and SEQ ID NO: 14, respectively;   (c) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 16, 17 and 18, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 20, GAS, and SEQ ID NO: 21, respectively;   (d) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 23, 24 and 25, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 27, GAS, and SEQ ID NO:28, respectively;   (e) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 30, 31 and 32, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 34, GAS, and SEQ ID NO: 35, respectively;   (f) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 37, 38 and 39, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 41, GAS, and SEQ ID NO: 42, respectively;   (g) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 6, GAS, and SEQ ID NO: 7, respectively;   (h) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 27, GAS, and SEQ ID NO: 28, respectively;   (i) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 34, GAS, and SEQ ID NO: 35, respectively;   (j) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 60, 61, and 62, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 13, the sequence DAS, and SEQ ID NO: 14;   (k) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 63, 64, and 65, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 20, the sequence GAS, and SEQ ID NO: 21; and   (l) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 66, 38, and 67, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 41, the sequence GAS, and SEQ ID NO: 42.   
     
     
         57 . The method of  claim 56 , wherein the antibody comprises a VH region and a VL region selected from the group consisting of:
 (a) a VH region comprising the sequence of SEQ ID NO: 1 and a VL region comprising the sequence of SEQ ID NO: 5;   (b) a VH region comprising the sequence of SEQ ID NO: 8 and a VL region comprising the sequence of SEQ ID NO: 12;   (c) a VH region comprising the sequence of SEQ ID NO: 15 and a VL region comprising the sequence of SEQ ID NO: 19;   (d) a VH region comprising the sequence of SEQ ID NO: 22 and a VL region comprising the sequence of SEQ ID NO: 26;   (e) a VH region comprising the sequence of SEQ ID NO: 29 and a VL region comprising the sequence of SEQ ID NO: 33;   (f) a VH region comprising the sequence of SEQ ID NO: 36 and a VL region comprising the sequence of SEQ ID NO: 40; and   (g) a variant of any of said antibodies, wherein said variant has at most 1, 2 or 3 amino acid substitutions.   
     
     
         58 . A method for treating cancer comprising tumor cells co-expressing HER2 and EGFR and/or HER3, the method comprising administering to an individual in need thereof an effective amount of an antibody comprising a VH region and a VL region selected from the group consisting of:
 (a) an antibody comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 2, 3 and 4, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 6, GAS, and SEQ ID NO: 7, respectively;   (b) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 9, 10 and 11, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 13, DAS, and SEQ ID NO: 14, respectively;   (c) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 16, 17 and 18, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 20, GAS, and SEQ ID NO: 21, respectively;   (d) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 23, 24 and 25, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 27, GAS, and SEQ ID NO: 28, respectively;   (e) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 30, 31 and 32, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 34, GAS, and SEQ ID NO: 35, respectively;   (f) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 37, 38 and 39, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 41, GAS, and SEQ ID NO: 42, respectively;   (g) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 6, GAS, and SEQ ID NO: 7, respectively;   (h) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 27, GAS, and SEQ ID NO: 28, respectively;   (i) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 34, GAS, and SEQ ID NO: 35, respectively;   (j) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 60, 61, and 62, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 13, the sequence DAS, and SEQ ID NO: 14;   (k) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 63, 64, and 65, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 20, the sequence GAS, and SEQ ID NO: 21; and   (l) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 66, 38, and 67, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 41, the sequence GAS, and SEQ ID NO: 42.   
     
     
         59 . The method of  claim 58 , wherein the antibody comprises a VH region and a VL region selected from the group consisting of:
 (a) a VH region comprising the sequence of SEQ ID NO: 1 and a VL region comprising the sequence of SEQ ID NO: 5;   (b) a VH region comprising the sequence of SEQ ID NO: 8 and a VL region comprising the sequence of SEQ ID NO: 12;   (c) a VH region comprising the sequence of SEQ ID NO: 15 and a VL region comprising the sequence of SEQ ID NO: 19;   (d) a VH region comprising the sequence of SEQ ID NO: 22 and a VL region comprising the sequence of SEQ ID NO: 26;   (e) a VH region comprising the sequence of SEQ ID NO: 29 and a VL region comprising the sequence of SEQ ID NO: 33;   (f) a VH region comprising the sequence of SEQ ID NO: 36 and a VL region comprising the sequence of SEQ ID NO: 40; and   (g) a variant of any of said antibodies, wherein said variant has at most 1, 2 or 3 amino acid substitutions.   
     
     
         60 . The method of  claim 58 , wherein the cancer is selected from the group consisting of breast cancer, colorectal cancer, endometrial/cervical cancer, lung cancer, malignant melanoma, ovarian cancer, pancreatic cancer, prostate cancer, testis cancer, a soft-tissue tumor such as synovial sarcoma, and bladder cancer. 
     
     
         61 . The method of  claim 58 , wherein the antibody is a bivalent antibody. 
     
     
         62 . The method of  claim 58 , wherein the antibody is an antigen-binding fragment. 
     
     
         63 . The method of  claim 58 , wherein the antibody is a full-length antibody. 
     
     
         64 . The method of  claim 58 , wherein the antibody is an effector-function-deficient antibody. 
     
     
         65 . The method of  claim 58 , wherein the antibody is a monovalent antibody. 
     
     
         66 . The method of  claim 65 , wherein the monovalent antibody further comprises a C H  region of an immunoglobulin or a fragment thereof comprising C H 2 and C H 3 regions, wherein the C H  region or fragment thereof has been modified such that the region corresponding to the hinge region and, if the immunoglobulin is not an IgG4 subtype, other regions of the C H  region do not comprise any amino acid residues which are capable of forming disulfide bonds with an identical C H  region or other covalent or stable non-covalent inter-heavy chain bonds with an identical C H  region in the presence of polyclonal human IgG. 
     
     
         67 . The method of  claim 58 , wherein the antibody is an IgG1 antibody. 
     
     
         68 . The method of  claim 67 , wherein the antibody is an IgG1,κ antibody. 
     
     
         69 . The antibody of  claim 58 , wherein the antibody is conjugated to another moiety. 
     
     
         70 . The antibody of  claim 69 , wherein the moiety is:
 (a) a cytotoxic moiety selected from the group consisting of taxol; cytochalasin B; gramicidin D; ethidium bromide; emetine; mitomycin; etoposide; tenoposide; vincristine; vinblastine; colchicin; doxorubicin; daunorubicin; dihydroxy anthracin dione; a tubulin-inhibitor; mitoxantrone; mithramycin; actinomycin D; 1-dehydrotestosterone; a glucocorticoid; procaine; tetracaine; lidocaine; propranolol; puromycin; calicheamicin or an analog or derivative thereof; an antimetabolite; an alkylating agent; an antibiotic; an antimitotic agent; diphtheria toxin; ricin toxin; cholera toxin; a Shiga-like toxin; LT toxin; C3 toxin; Shiga toxin; pertussis toxin; tetanus toxin; soybean Bowman-Birk protease inhibitor;  Pseudomonas  exotoxin; alorin; saporin; modeccin; gelanin; abrin A chain; modeccin A chain; alpha-sarcin;  Aleurites fordii  proteins; dianthin proteins;  Phytolacca americana  proteins;  Momordica charantia  inhibitor; curcin; crotin;  Sapaonaria officinalis  inhibitor; gelonin; mitogellin; restrictocin; phenomycin; enomycin toxins; ribonuclease (RNase); DNase I; Staphylococcal enterotoxin A; pokeweed antiviral protein; diphtherin toxin; and  Pseudomonas  endotoxin;   (b) a cytotoxic moiety selected from the group consisting of maytansine, calicheamicin, duocarmycin, rachelmycin (CC-1065), monomethyl auristatin E, and an analog, derivative, or prodrug of any thereof;   (c) a cytokine selected from the group consisting of interleukin (IL)-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, keratinocyte growth factor, interferon (IFN)α, IFNβ, IFNγ, granulocyte-macrophage colony-stimulating factor, CD40L, Flt3 ligand, stem cell factor, ancestim, and tumor necrosis factor (TNF)α; or   (d) a radioisotope.   
     
     
         71 . The method of  claim 58 , wherein the antibody is a bispecific antibody comprising a second antigen-binding site. 
     
     
         72 . The method of  claim 71 , wherein the second antigen-binding site binds to a molecule selected from the group consisting of: a cancer- or tumor-associated antigen; a cancer-associated integrin; a T cell and/or NK cell antigen; an angiogenic factor, receptor for an angiogenic factor, or other cancer-associated growth factor; and a receptor associated with cancer progression. 
     
     
         73 . The method of  claim 72 , wherein the cancer- or tumor-associated antigen is selected from the group consisting of: a carcinoembryonic antigen, prostate specific antigen, renal antigen, α-fetoprotein, CTL-recognized antigen on melanoma, a CT antigen, a mucin antigen, a ganglioside antigen, tyrosinase, gp75, c-Met, c-myc, Mart1, MelanA, MUM-1, MUM-2, MUM-3, HLA-B7, and Ep-CAM. 
     
     
         74 . The method of  claim 73 , wherein:
 (a) the CT antigen is selected from the group consisting of: MAGE-B5, MAGE-B6, MAGE-C2, MAGE-C3, MAGE-D, Mage-12, CT10, NY-ESO-1, SSX-2, GAGE, BAGE, MAGE, and SAGE;   (b) the mucin antigen is MUC1 or mucin-CA125;   (c) the cancer-associated integrin is α5β3 integrin;   (d) the T cell and/or NK cell antigen is CD3 or CD16;   (e) the angiogenic factor or other cancer-associated growth factor is selected from the group consisting of vascular endothelial growth factor, fibroblast growth factor, epidermal growth factor, angiogenin, vascular endothelial growth factor receptor, fibroblast growth factor receptor, epidermal growth factor receptor, and angiogenin receptor; or   (f) the receptor associated with cancer progression is selected from the group consisting of HER1, HER3, and HER4.   
     
     
         75 . A method for detecting the presence of human epidermal growth factor receptor 2 (HER2) in a sample, comprising:
 (a) contacting the sample with an antibody which binds to HER2 under conditions that allow for formation of a complex between the antibody and HER2, wherein the antibody comprises a VH region and a VL region selected from the group consisting of:
 (i) an antibody comprising a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 2, 3 and 4, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 6, GAS, and SEQ ID NO: 7, respectively; 
 (ii) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 9, 10 and 11, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 13, DAS, and SEQ ID NO: 14, respectively; 
 (iii) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 16, 17 and 18, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 20, GAS, and SEQ ID NO: 21, respectively; 
 (iv) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 23, 24 and 25, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 27, GAS, and SEQ ID NO: 28, respectively; 
 (v) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 30, 31 and 32, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 34, GAS, and SEQ ID NO: 35, respectively; 
 (vi) a VH region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs: 37, 38 and 39, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 41, GAS, and SEQ ID NO: 42, respectively; 
 (vii) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 6, GAS, and SEQ ID NO: 7, respectively; 
 (viii) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 27, GAS, and SEQ ID NO: 28, respectively; 
 (ix) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 34, GAS, and SEQ ID NO: 35, respectively; 
 (x) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 60, 61, and 62, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 13, the sequence DAS, and SEQ ID NO: 14; 
 (xi) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 63, 64, and 65, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 20, the sequence GAS, and SEQ ID NO: 21; and 
 (xii) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 66, 38, and 67, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NO: 41, the sequence GAS, and SEQ ID NO: 42; and 
   (b) analyzing whether a complex has been formed.