Production of enzymatic active glucocerebrosidase of recombinant cells.

摘要

A method is provided for the production of biologically active glucocerebrosidase (GCR) in a cell culture. In one embodiment, recombinant cells containing the DNA sequence encoding for the production of GCR are cultured in a growth medium suitable for the enhancement of cell growth. A substantial portion of the growth medium may then be replaced with a production medium that is suitable for cell viability and secretion and stabilization of GCR. After production and secretion of GCR into the production medium, the production medium may be removed and collected. The GCR may then be purified from the collected production medium. Compositions, that are not toxic to the cells, may be added to the production medium to stabilize the secreted GCR. Stabilizing compositions that may be added include thiol-containing compositions such as dithiothreitol, glutathione, 1,3-mercaptoglycerol, cysteamine, lipoic acid, lipoamide and thioglycolate or proteins such as serum or serum albumin. In addition, the pH or dissolved oxygen concentration of the production medium may be adjusted, or a combination of conditions may be utilized.