HUMAN ANTIBODIES THAT BIND HUMAN TNF-ALPHA AND METHODS OF PREPARING THE SAME
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.
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.