Methods to manipulate quality attributes of polypeptides produced in cho cells
AssigneeBristol-Myers Squibb Company
InventorJun Tian
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Abstract
In accordance with the present invention, CHO cells expressing a recombinant polypeptide of interest are grown in media where the amino acids, vitamins, phosphate, lipids and/or antioxidant optimization is utilized to manipulate and/or control the protein quality attributes of the polypeptides. Polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical compositions.
Claims12
A method of increasing galactose content of a recombinant glycoprotein expressed in Chinese Hamster Ovary (CHO) cells under suitable cell culture conditions comprising:
- adding at least about 100 mg/ml of zinc and less than about 0.1 g/L asparagine 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 glycoprotein is increased.
A method of decreasing galactose content of a recombinant glycoprotein expressed in Chinese Hamster Ovary (CHO) cells under suitable cell culture conditions comprising:
- 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 glycoprotein is increased.
Dependencies are inferred from the claim text.