Human antibodies that bind human tnf-alpha and methods of preparing the same

Abstract

Methylglyoxal (MGO)-modified recombinant TNF-alpha antibodies (e.g., Adalimumab) are identified. MGO modification decreases binding between Adalimumab and TNF-alpha. Methods are disclosed for reducing the presence of MGO-modified antibodies in the production of Adalimumab TNF-alpha antibodies.

We claim: 
     
         1 . A composition comprising a binding protein capable of binding TNF-alpha, said binding protein comprising a methylglyoxal (MGO)-susceptible amino acid, wherein said composition is prepared by substantially removing molecules of said binding protein that comprise at least one MGO-modified amino acid. 
     
     
         2 . The composition of  claim 1 , wherein more than 70% of said molecules that comprise at least one MGO-modified amino acid is removed. 
     
     
         3 . The composition of  claim 1 , wherein more than 90% of said molecules that comprise at least one MGO-modified amino acid is removed. 
     
     
         4 . The composition of  claim 1 , wherein the MGO-susceptible amino acid is an arginine. 
     
     
         5 . The composition of  claim 1 , wherein the binding protein is a human antibody or an antigen-binding portion thereof, wherein the binding protein dissociates from human TNF-alpha with a K d  of 1×10 −8  M or less and a K off  rate constant of 1×10 −3  s −1  or less, both determined by surface plasmon resonance, and wherein the binding protein neutralizes human TNF-alpha cytotoxicity in a standard in vitro L929 assay with an IC 50  of 1×10 −7  M or less.