Methods of preventing and removing trisulfide bonds
AssigneeBiogen MA Inc.
InventorsRashmi Rohit Kshirsagar, Alan Gilbert
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Abstract
The present invention pertains to methods of preventing and eliminating trisulfide bonds in proteins such as antibodies. In one embodiment, trisulfide bonds in proteins are converted to disulfide bonds as part of chromatographic purification procedures. In another embodiment, the formation of trisulfide bonds in proteins is inhibited by implementation of methods described herein during the cell culture production of such proteins. In another embodiment, monoclonal antibodies are produced by the methods described herein.
Claims13
A method for reducing the formation of trisulfide bonds in proteins during large-scale production comprising culturing cells expressing said proteins in the presence of an effective amount of an inhibitor of cysteine degradation, whereby trisulfide linkage formation in said proteins is reduced relative to cells cultured in medium without the inhibitor of cysteine degradation, wherein the inhibitor of cysteine degradation is formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, stearic acid, arachidonic acid, docosahexaenoic acid, myristoleic acid, palmitoleic acid, elaidic acid, erucic acid, vaccenic acid, penicillamine, carotenes, alpha-tocopherol, ubiquinol, lactic acid, formic acid, oxalic acid, or uric acid, and wherein said cells are mammalian cells.
Dependencies are inferred from the claim text.