Methods of minimizing glycoprotein agregation in CHO cell production
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.
1. A method of decreasing aggregation of a recombinant glycoprotein expressed in Chinese Hamster Ovary (CHO) cells under suitable cell culture conditions comprising:
adding 1 mg/L-100 mg/L of Zinc, 1.05 g/L-2 g/L Asparagine and 0.1 g-1.05 g/L Aspartic 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
and wherein the level of aggregation of the expressed glycoprotein is decreased.
2. The method of claim 1 , wherein the cell culture medium is chemically defined.
3. The method of claim 2 , wherein the chemically defined cell culture medium is basal medium.
4. The method of claim 3 , wherein the chemically defined basal medium does not contain added serum or hydrolysates.
5. The method of claim 3 , wherein the chemically defined basal medium is protein-free.
6. The method according to claim 1 , wherein the glycoprotein is a recombinant antibody, antibody fragment or fusion protein.
7. The method according to claim 1 , wherein the cell culture process is performed under batch, fed-batch or continuous cell culture conditions.