Patent ID: 8129145

Claim:
A method of producing a glycoprotein in a cell culture comprising: culturing mammalian cells that contain a gene encoding a glycoprotein of interest in a cell culture medium comprising between approximately 10 nM and 600 nM manganese, wherein the glycoprotein of interest comprises O-linked oligosaccharide chains; maintaining the culture at a first temperature range of about 30 to 42 degrees Celsius for a first period of time sufficient to allow the cells to reproduce to a viable cell density within a range of about 20%-80% of the maximal possible viable cell density if the culture were maintained at the first temperature range; and shifting the culture to a second temperature range and maintaining the culture for a second period of time sufficient to permit expression of the glycoprotein, wherein the second temperature range comprises a temperature range that is approximately 25 to 41 degrees Celsius, wherein the glycosylation pattern of the expressed glycoprotein is more extensive than the glycosylation pattern observed under otherwise identical conditions in otherwise identical medium that lacks the manganese.