Antigen associated with rheumatoid arthritis

Abstract

The invention relates to a binding member that binds the Extra Domain-A (ED-A) isoform of fibronectin for the detection and treatment of rheumatoid arthritis.

What is claimed: 
     
         1 . An isolated nucleic acid molecule encoding an antibody conjugate, wherein the antibody conjugate comprises an antibody, or an antigen-binding fragment thereof that binds the Extra Domain-A (ED-A) of fibronectin conjugated to a molecule having anti-inflammatory activity, wherein
 said antibody comprises a heavy chain variable (VH) domain and a light chain variable (VL) domain,
 said VH domain comprises a set of complementarity determining regions HCDR1, HCDR2 and HCDR3 that comprise the amino acid sequences of SEQ ID NOs: 83, 4, and 5, respectively, and 
 said VL domain comprises a set of complementarity determining regions LCDR1, LCDR2 and LCDR3 that comprise the amino acid sequences of SEQ ID NOs: 86, 7, and 8, respectively. 
   
     
     
         2 . The isolated nucleic acid molecule according to  claim 1 , wherein:
 i) said VH domain comprises a human framework;   ii) said VL domain comprises a human framework; or   iii) said VH domain comprises a human framework and said VL domain comprises a human framework.   
     
     
         3 . The isolated nucleic acid molecule according to  claim 1 , wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 81; or the amino acid sequence of SEQ ID NO: 81, wherein the amino acid at position 5 of SEQ ID NO: 81 is a leucine residue (L) rather than a valine residue (V). 
     
     
         4 . The isolated nucleic acid molecule according to  claim 1 , wherein the VL domain comprises the amino acid sequence of SEQ ID NO: 82; or the amino acid sequence of SEQ ID NO: 82, wherein the amino acid at position 18 of SEQ ID NO: 82 is an arginine residue (R) rather than a lysine residue (K). 
     
     
         5 . The isolated nucleic acid molecule according to  claim 1 , wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 81, wherein the amino acid at position 5 of SEQ ID NO: 81 is a leucine residue (L) rather than a valine residue (V); and the VL domain comprises the amino acid sequence of SEQ ID NO: 82, wherein the amino acid at position 18 of SEQ ID NO: 82 is an arginine residue (R) rather than a lysine residue (K). 
     
     
         6 . The isolated nucleic acid molecule according to  claim 5 , wherein said antigen-binding fragment comprises a single chain Fv (scFv) or is a diabody. 
     
     
         7 . The isolated nucleic acid molecule according to  claim 6 , wherein said molecule having anti-inflammatory activity is human interleukin-10 (IL-10). 
     
     
         8 . The isolated nucleic acid molecule according to  claim 7 , wherein said human IL-10 comprises amino acid residues 258 to 417 of SEQ ID NO: 149. 
     
     
         9 . The isolated nucleic acid molecule according to  claim 5 , wherein said antibody or antigen-binding fragment is conjugated to said molecule having anti-inflammatory activity via a peptide linker. 
     
     
         10 . The isolated nucleic acid molecule according to  claim 9 , wherein said peptide linker comprises 15 amino acids. 
     
     
         11 . The isolated nucleic acid molecule according to  claim 10 , wherein said peptide linker comprises the amino acid sequence (SSSSG) 3  (amino acid residues 243-257 of SEQ ID NO: 149). 
     
     
         12 . The isolated nucleic acid molecule according to  claim 5 , wherein said antigen-binding fragment comprises a scFv or is a diabody, and wherein said VH domain is conjugated to said VL domain via an amino acid linker. 
     
     
         13 . The isolated nucleic acid molecule according to  claim 12 , wherein said amino acid linker comprises 5 to 25 amino acids. 
     
     
         14 . The isolated nucleic acid molecule according to  claim 12 , wherein said amino acid linker comprises 5 amino acids. 
     
     
         15 . The isolated nucleic acid molecule according to  claim 1 , wherein the antibody conjugate comprises:
 (i) the antigen-binding fragment, wherein the antigen-binding fragment comprises a scFv or is a diabody comprising said VH domain and said VL domain,   wherein said VH domain is conjugated to said VL domain via a 5-amino acid linker; and   (ii) human interleukin-10,   wherein said VL domain is conjugated to said human interleukin-10 via a peptide linker comprising the amino acid sequence (SSSSG) 3  (amino acid residues 243-257 of SEQ ID NO: 149).   
     
     
         16 . The isolated nucleic acid molecule according to  claim 1 , wherein the antibody conjugate consists of:
 (i) the antigen-binding fragment, wherein the antigen-binding fragment is a diabody consisting of:
 (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 81, wherein the amino acid at position 5 of SEQ ID NO: 81 is a leucine residue (L) rather than a valine residue (V); and 
 (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 82, wherein the amino acid at position 18 of SEQ ID NO: 82 is an arginine residue (R) rather than a lysine residue (K); 
   (ii) a 5-amino acid linker consisting of the amino acid sequence, wherein said VH domain is conjugated to said VL domain via the 5-amino acid linker;   (iii) human interleukin-10; wherein said human IL-10 comprises amino acid residues 258 to 417 of SEQ ID NO: 149, and   (iv) a peptide linker consisting of the amino acid sequence (SSSSG) 3  (amino acid residues 243-257 of SEQ ID NO: 149), wherein said VL domain is conjugated to said human interleukin-10 via the peptide linker.   
     
     
         17 . The isolated nucleic acid molecule according to  claim 2 , wherein:
 i) said VH domain comprises a framework from the human germline DP47 gene;   ii) said VL domain comprises a framework from the human germline DPK22 gene; or   iii) VH domain comprises a framework from the human germline DP47 gene and said VL domain comprises a framework from the human germline DPK22 gene.   
     
     
         18 . A host cell comprising the isolated nucleic acid molecule according to  claim 1 . 
     
     
         19 . A method of producing an antibody conjugate comprising an antibody, or an antigen-binding fragment thereof that binds the Extra Domain-A (ED-A) of fibronectin conjugated to a molecule having anti-inflammatory activity, wherein the method comprises culturing the host cell of  claim 18  under conditions suitable for production of said antibody conjugate. 
     
     
         20 . The method according to  claim 19 , further comprising isolating said antibody conjugate.