	US 12,234,483 B2
	Methods of decreasing the galactose content of a recombinant glycoprotein expressed in CHO cells
Jun Tian, Westford, MA (US)
Assigned to Bristol-Myers Squibb Company, Princeton, NJ (US)
Filed by BRISTOL-MYERS SQUIBB COMPANY, Princeton, NJ (US)
Filed on Apr. 18, 2023, as Appl. No. 18/302,043.
Application 18/302,043 is a division of application No. 17/830,794, filed on Jun. 2, 2022, granted, now 11,655,457.
Application 17/830,794 is a division of application No. 15/774,138, granted, now 11,401,509, issued on Aug. 2, 2022, previously published as PCT/US2016/060782, filed on Nov. 7, 2016.
Claims priority of provisional application 62/252,849, filed on Nov. 9, 2015.
Prior Publication US 2023/0250399 A1, Aug. 10, 2023
Int. Cl. C12N 5/071 (2010.01); C12N 5/00 (2006.01); C12P 21/00 (2006.01)

CPC C12N 5/0682 (2013.01) [C12N 5/0031 (2013.01); C12P 21/005 (2013.01); C12N 2500/22 (2013.01); C12N 2500/24 (2013.01); C12N 2500/32 (2013.01); C12N 2510/02 (2013.01)]

6 Claims

1. A method of decreasing galactose content of a recombinant glycoprotein comprising:

culturing a Chinese Hamster Ovary (CHO) host cell in a suitable cell culture medium, wherein the CHO host cell comprises an expression vector encoding a glycoprotein;

adding at least about 2 g/L asparagine and less than about 0.01 mg/L zinc to the cell culture medium,

wherein the cell culture medium is maintained under following conditions:

a) pH between 6.5-7.5;

b) dissolved oxygen between 5% and 90% of air saturation;

c) carbon dioxide between 10 mmHg and 150 mmHg; and

d) temperature between 30° C. and 37° C.; and

wherein the galactose content of the expressed glycoprotein is decreased.