Process for modifying galactosylation and G0F content of a glycoprotein composition by glutamine supplementation
AssigneeDr. Reddy's Laboratories, Inc.
InventorsSreenath Kadreppa, Madhava Ram Paranandi Ananta, Senthil Kumar Baskaran, Shinto Antony
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Abstract
The invention relates to a cell culture process for decreasing the galactosylated content and/or increasing the G0F content of a glycoprotein. The process involves subjecting recombinant cells expressing the said glycoprotein to a temperature and pH shift and supplementing cell culture with glutamine.
Claims15
A cell culture process for obtaining an anti-VEGF antibody composition comprising the steps of,
- culturing Chinese Hamster Ovary (CHO) cells at a first temperature and a first pH for a first period of time,
- shifting the temperature and the pH to a second value, where the second temperature value is lower than the first temperature and the second pH value is lower than the first pH, and
- supplementing the said culture with glutamine,
- wherein the concentration of glutamine in the culture is about 4 mM to about 12 mM, and
- wherein the change in temperature and pH is done simultaneously at about two days to about three days after the inoculation of cell culture, thereby obtaining the antibody composition comprising of about 16% to about 25% galactosylated glycan and/or about 66% to about 74% G0F glycan.
A cell culture process for increasing the G0F glycans in an anti-VEGF antibody composition, the process comprising the steps of,
- culturing Chinese Hamster Ovary (CHO) cells at first temperature and first pH for a first period of time,
- shifting the temperature and the pH to a second value, where the second value is lower than the first temperature and first pH,
- supplementing the said culture with glutamine,
- wherein the concentration of glutamine in the culture is about 4 mM to about 12 mM, and
- wherein the change in temperature and pH is done simultaneously at about two days to about three days after the inoculation of cell culture and wherein the increase is of about 13% to about 21% when compared to an antibody composition obtained by a process without addition of glutamine.
A cell culture process for decreasing the galactosylated glycans in an anti-VEGF antibody composition the process comprising the steps of,
- culturing Chinese Hamster Ovary (CHO) cells at first temperature and first pH for a first period of time,
- shifting the temperature and the pH to a second value, where the second value is lower than the first temperature and first pH
- supplementing the said culture with glutamine,
- wherein the concentration of glutamine in the culture is about 4 mM to about 12 mM, and
- wherein the change in temperature and pH is done simultaneously at about two days to about three days after the inoculation of cell culture and wherein the decrease is of about 27% to about 46% when compared to an antibody composition obtained by a process without addition of glutamine.
Dependencies are inferred from the claim text.