Dual Variable Domain Immunoglobulin and Uses Thereof

Abstract

The present invention relates to engineered multivalent and multispecific binding proteins, methods of making, and specifically to their uses in the prevention and/or treatment of acute and chronic inflammatory and other diseases.

1 - 65 . (canceled) 
     
     
         66 . A method of producing a binding protein, comprising culturing a host cell in culture medium, wherein said host cell comprises:
 (a) a first nucleic acid encoding a first polypeptide chain of the binding protein that comprises formula 1: VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, C is a heavy chain constant domain, X1 is a linker with the proviso that it is not CH1, X2 is an Fc region, and n is 0 or 1; and   (b) a second nucleic acid encoding a second polypeptide chain of the binding protein that comprises formula 2: VD1-(X1)n-VD2-C, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain, C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and n is 0 or 1;   wherein the first nucleic acid is present on the same or a different expression vector as the second nucleic acid;   wherein the culturing of the host cell is under conditions sufficient to produce the first and second polypeptide chains; and   wherein one of the first polypeptide chain and one of the second polypeptide chain form a divalent form of the binding protein comprising two antigen binding sites, and wherein two of the first polypeptide chains and two of the second polypeptides form a tetravalent form of the binding protein comprising four antigen binding sites.   
     
     
         67 . The method of  claim 66 , wherein 50%-75% of the binding protein produced is a dual specific tetravalent binding protein. 
     
     
         68 . The method of  claim 66 , wherein 75%-90% of the binding protein produced is a dual specific tetravalent binding protein. 
     
     
         69 . The method of  claim 66 , wherein 90%-95% of the binding protein produced is a dual specific tetravalent binding protein. 
     
     
         70 - 76 . (canceled)