Control of Protein Glycosylation by Culture Medium Supplementation and Cell Culture Process Parameters

A method of altering the glycosylation pattern of a recombinant glycoprotein produced in cell culture comprising:

• culturing eukaryotic cells engineered to express a recombinant glycoprotein of interest in a cell culture medium, wherein the cell culture medium is supplemented with an additive comprising one or more of mycophenolic acid, mycophenolic acid acyl glucuronide, insulin, copper (II) sulfate, hypoxanthine, thymidine, guanine, glucosamine, or galactose;
• wherein the glycosylation pattern of the recombinant glycoprotein of interest is altered to better resemble the glycosylation pattern of a reference sample of the glycoprotein.

A method of altering the glycosylation pattern of a recombinant glycoprotein produced in cell culture comprising:

• supplementing the culture medium of a cell culture of eukaryotic cells engineered to express a recombinant glycoprotein of interest with an additive comprising one or more of mycophenolic acid, mycophenolic acid acyl glucuronide, insulin, copper (II) sulfate, hypoxanthine, thymidine, guanine, glucosamine, or galactose;
• wherein the glycosylation pattern of the recombinant glycoprotein of interest is altered relative to the same recombinant glycoprotein produced by the same cells in the same cell culture medium without the additive.

A method of altering the glycosylation pattern of a recombinant glycoprotein produced in cell culture comprising:

• supplementing the culture medium of a cell culture of eukaryotic cells engineered to express a recombinant glycoprotein of interest with an additive comprising one or more of mycophenolic acid, mycophenolic acid acyl glucuronide, insulin, copper (II) sulfate, hypoxanthine, thymidine, guanine, glucosamine, or galactose;
• wherein the glycosylation pattern of the recombinant glycoprotein of interest is altered to better resemble the glycosylation pattern of a reference sample of the glycoprotein.

15-172. (canceled)