Prevention of disulfide bond reduction during recombinant production of polypeptides
AssigneeGenentech, Inc.
InventorsYung-Hsiang Kao, Michael W. Laird, Melody Trexler Schmidt, Rita L. Wong, Daniel P. Hewitt
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Abstract
The invention concerns methods for preventing the reduction of disulfide bonds during the recombinant production of disulfide-containing polypeptides. In particular, the invention concerns the prevention of disulfide bond reduction during harvesting of disulfide-containing polypeptides, including antibodies, from recombinant host cell cultures.
Claims43
A method for the prevention of the reduction of a disulfide bond in a therapeutic IgG1 antibody expressed by a recombinant Chinese Hamster Ovary (CHO) host cell, comprising, following fermentation, lowering the pH of the harvested cell culture fluid (HCCF) of the recombinant CHO host cell, wherein lowering the pH of the HCCF results in prevention of the reduction of the disulfide bond in the antibody, and wherein the pH is lowered to a value of less than pH 6.
A method for the prevention of the reduction of a disulfide bond in a therapeutic IgG1 antibody expressed by a recombinant Chinese Hamster Ovary (CHO) host cell, comprising, following fermentation and immediately after harvest, lowering the pH of the harvested cell culture fluid (HCCF) of the recombinant CHO host cell, wherein lowering the pH of the HCCF results in prevention of the reduction of the disulfide bond in the therapeutic IgG1 antibody, and wherein the pH is lowered to a value of less than pH 6.
A method for the prevention of the reduction of a disulfide bond in a therapeutic protein comprising an Fc region of an IgG1 antibody expressed by a recombinant Chinese Hamster Ovary (CHO) host cell, comprising, following fermentation, lowering the pH of the harvested cell culture fluid (HCCF) of the recombinant CHO host cell, wherein lowering the pH of the HCCF results in prevention of the reduction of the disulfide bond in the therapeutic protein, wherein the pH is lowered to a value of less than pH 6.
Dependencies are inferred from the claim text.