Method of producing recombinant high molecular weight vwf in cell culture

摘要

Discloses a method of producing a recombinant Von Willebrand Factor (rVWF) composition, the method comprising; (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 l.Lg/L; (c) providing one or more cells comprising a nucleic acid encoding a rVWF protein; (d) culturing the one or more cells in the cell culture media supplemented with copper such that rVWF is expressed and excreted from the cells into a culture supernatant and the NH4 + content of the culture supernatant is maintained at a concentration below 4 mM or 10 mM; and (e) recovering at least a portion of the culture supernatant, wherein the supernatant recovered has a rVWF specific ristocetin cofactor activity of at least 30 mU/l.Lg rVWF