Method of producing recombinant high molecular weight vWF in cell culture
AssigneesBaxter International Inc., Baxter Healthcare S.A.
InventorsLeopold Grillberger, Manfred Reiter, Wolfgang Mundt
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Abstract
Among other aspects, the present invention relates to cell culture conditions for producing high molecular weight vWF, in particular, highly multimericWF with a high specific activity and ADAMTS13 with a high specific activity. The cell culture conditions of the present invention can include, for example, a cell culture medium with an increased copper concentration and/or cell culture supernatant with a low ammonium (NH4+) concentration. The present invention also provides methods for cultivating cells in the cell culture conditions to express high molecular weight vWF and rA13 having high specific activities.
Claims35
A method for producing a recombinant Von Willebrand Factor (rVWF) composition, the method comprising the steps of:
- (a) providing a basal cell culture media;
- (b) supplementing the basal cell culture media with copper to provide a final copper concentration of at least 2.4 μg/L;
- (c) providing one or more cells comprising a nucleic acid encoding a rVWF protein;
- (d) culturing the one or more cells in the copper supplemented cell culture media such that rVWF is expressed and excreted from the cells into a culture supernatant and the NH4+content of the cell culture supernatant is maintained at a concentration below 4 mM or below 10 mM;
- and
- (e) recovering at least a portion of the culture supernatant,
- wherein the recovered supernatant has a rVWF specific ristocetin cofactor activity of at least 30 mU/μg rVWF.
Dependencies are inferred from the claim text.