HUMAN ANTIBODIES THAT BIND HUMAN TNFalpha

Abstract

Human antibodies, preferably recombinant human antibodies, that specifically bind to human tumor necrosis factor α (hTNFα) are disclosed. These antibodies have high affinity for hTNFα (e.g., K d =10 −8 M or less), a slow off rate for hTNFα dissociation (e.g., K off =10 −3 sec −1 or less) and neutralize hTNFα activity in vitro and in vivo. An antibody of the invention can be a full-length antibody or an antigen-binding portion thereof. The antibodies, or antibody portions, of the invention are useful for detecting hTNFα and for inhibiting hTNFα activity, e.g., in a human subject suffering from a disorder in which hTNFα activity is detrimental. Nucleic acids, vectors and host cells for expressing the recombinant human antibodies of the invention, and methods of synthesizing the recombinant human antibodies, are also encompassed by the invention.

1 . An isolated human antibody, or an antigen-binding portion thereof, which dissociates from human TNFα with a Kd of 1×10 −8  M or less and a Koff rate constant of 1×10 −3  s −1  or less, both determined by surface plasmon resonance, and neutralizes TNFα-induced cellular activation in a standard in vitro assay for TNFα-induced ELAM-1 expression on human umbilical vein endothelial cells (HUVEC). 
     
     
         2 . The isolated human antibody of  claim 1 , wherein the isolated human antibody, or antigen-binding portion thereof, comprises
 a light chain variable region (LCVR) comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3, or modified from SEQ ID NO: 3 by a single alanine substitution at position 1, 4, 5, 7 or 8 or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 7; and   comprises a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to five conservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 8.   
     
     
         3 . The isolated human antibody, or an antigen-binding portion thereof, of  claim 1 , wherein the antibody has a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 1 and a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2. 
     
     
         4 . The isolated human antibody, or an antigen-binding portion thereof, of  claim 1 , wherein the antibody has an IgG1 heavy chain constant region. 
     
     
         5 . The isolated human antibody, or an antigen-binding portion thereof, of  claim 1 , wherein the antibody has a kappa light chain constant region. 
     
     
         6 . The isolated human antibody, or an antigen-binding portion thereof, of  claim 1 , wherein the antibody is a Fab fragment or a single chain Fv fragment. 
     
     
         7 . A method of treating a subject having a disorder in which TNFα is detrimental, comprising administering an isolated human antibody, or an antigen-binding portion thereof, which dissociates from human TNFα with a Kd of 1×10 −8  M or less and a Koff rate constant of 1×10 −3  s −1  or less, both determined by surface plasmon resonance, and neutralizes TNFα-induced cellular activation in a standard in vitro assay for TNFα-induced ELAM-1 expression on human umbilical vein endothelial cells (HUVEC), such that the disorder in which TNFα is detrimental is treated. 
     
     
         8 . The method of  claim 7 , wherein the disorder is an autoimmune disease. 
     
     
         9 . The method of  claim 8 , wherein the autoimmune disease is selected from the group consisting of: rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis, an allergy, multiple sclerosis, autoimmune diabetes, autoimmune uveitis and nephrotic syndrome. 
     
     
         10 . The method of  claim 7 , wherein the disorder is an infectious disease. 
     
     
         11 . The method of  claim 7 , wherein the disorder is transplant rejection or graft-versus-host disease. 
     
     
         12 . The method of  claim 7 , wherein the disorder is a malignancy. 
     
     
         13 . The method of  claim 7 , wherein the disorder is a pulmonary disorder. 
     
     
         14 . The method of  claim 7 , wherein the disorder is an intestinal disorder. 
     
     
         15 . The method of  claim 14 , wherein the intestinal disorder is Crohn's disease or ulcerative colitis. 
     
     
         16 . The method of  claim 7 , wherein the disorder is a cardiac disorder. 
     
     
         17 . The method of  claim 7 , wherein the disorder is selected from the group consisting of inflammatory bone disorders, bone resorption disease, alcoholic hepatitis, viral hepatitis, coagulation disturbances, burns, reperfusion injury, keloid formation, scar tissue formation and pyrexia. 
     
     
         18 . The method of  claim 1 , wherein the isolated human antibody, or antigen-binding portion thereof, comprises
 a light chain variable region (LCVR) comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3, or modified from SEQ ID NO: 3 by a single alanine substitution at position 1, 4, 5, 7 or 8 or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 7; and   comprises a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to five conservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 8.   
     
     
         19 . The method of  claim 7 , wherein the antibody has a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 1 and a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2.