Human antibodies that bind human tnf-alpha and methods of preparing the same
AssigneeAbbVie Inc.
InventorChris M. CHUMSAE
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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 MOO-modified antibodies in the production of Adalimumab TNF-alpha antibodies.
Claims21
A composition comprising a binding protein capable of binding TNF-alpha, wherein said binding protein comprises at least one methylglyoxal (MGO)-susceptible amino acid, and wherein at least a portion of said binding protein comprises one or more MGO-modified amino acids.
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.
A method for purifying a composition comprising a target protein, said method comprising:
- (a) loading the composition to a cation exchange adsorbent using a loading buffer, wherein the pH of the loading buffer is lower than the pI of the target protein;
- (b) washing the cation exchange adsorbent with a washing buffer, wherein the pH of the washing buffer is lower than the pI of the target protein;
- (c) eluting the cation exchange adsorbent with an elution buffer, said elution buffer being capable of reducing the binding between the target protein and the cation exchange adsorbent;
- and
- (d) collecting the eluate, wherein the percentage of the target protein is higher in the eluate than the percentage of the target protein in the composition.
A method for purifying a composition comprising a target protein, said method comprising:
- (a) loading the composition to an anion exchange adsorbent using a loading buffer, wherein the pH of the loading buffer is lower than the isoelectric point (pI) of the target protein;
- (b) allowing the majority of the target protein to pass through without binding to the anion exchange adsorbent;
- (c) collecting the pass-through loading buffer containing said unbound target protein;
- (d) washing the anion exchange adsorbent with a washing buffer;
- (e) allowing the target protein bound to the anion exchange adsorbent to disassociate from the anion exchange adsorbent;
- and
- (f) collecting the washing buffer containing said disassociated target protein.
Dependencies are inferred from the claim text.