Process for improving Omalizumab expression in CHO cells
The present invention relates to a process of improving the production of anti-IgE antibody specifically Omalizumab with improved titer and reduced galactosylation and/or reduced acidic variant expressed in mammalian cell culture in suitable culture condition for an example using fed batch mode by maintaining specific temperature 36.5° C.±0.5 throughout the duration of cells culture.
1 . A process for improving the expression of omalizumab in Chinese Hamster Ovary cell culture comprising;
a) culturing the Chinese Hamster Ovary cells capable to express omalizumab in the Chinese Hamster Ovary cell culture;
b) maintaining suitable temperature at 36.5° C. throughout the Chinese Hamster Ovary cell culture;
c) harvesting the omalizumab from the Chinese Hamster Ovary cell culture,
wherein the process improves titer of the omalizumab compared to cell culture process performed at temperature 35.5° C. or 37.5° C., and
wherein the process reduces total galactosylation of the omalizumab or reduces expression of acidic variants of the omalizumab compared to cell culture process performed at temperature 35.5° C. or 37.5° C.
2 . The process as claimed in claim 1 , wherein the Chinese Hamster Ovary culture comprises a growth phase and a production phase, wherein the production phase maintains suitable pH selected from pH 6.7, pH 6.8, and pH 6.9.
3 . The process as claimed in claim 1 , wherein the process increases the titer of the omalizumab by 50%±5% to 190±19% compared to cell culture process performed at temperature 35.5° C. or 37.5° C.
4 . The process as claimed in claim 1 , wherein the process reduces total galactosylation of the omalizumab 2%±0.2% to 50%±5% compared to cell culture process performed at temperature 35.5° C. or 37.5° C.
5 . The process as claimed in claim 1 , wherein the process results in total galactosylation of the omalizumab of less than 13%.
6 . The process as claimed in claim 1 , wherein the process results in total galactosylation of the omalizumab of 8%±0.8% to 12%±1.2%.
7 . The process as claimed in claim 1 , wherein the process reduces expression of acidic variants of the omalizumab 5%±0.5% to 50%±5% compared to cell culture process performed at temperature 35.5° C. or 37.5° C.
8 . The process as claimed in claim 1 , wherein the process results in expression of acidic variants of the omalizumab of less than 17%.
9 . The process as claimed in claim 1 , wherein the process results in expression of acidic variants of the omalizumab of 12%±1.2% to 16%±1.6% compared to cell culture process performed at temperature 35.5° C. or 37.5° C.
10 . A process for improving the expression of omalizumab in Chinese Hamster Ovary cell culture comprising:
a) culturing the Chinese Hamster Ovary cells capable to express omalizumab in the Chinese Hamster Ovary cell culture;
b) maintaining temperature 36.5° C. throughout the Chinese Hamster Ovary cell culture;
c) maintaining pH 6.8±0.1 during the production phase of mammalian cell culture;
d) harvesting omalizumab from Chinese Hamster Ovary cell culture,
wherein the process improves titer of the omalizumab compared to cell culture process performed at temperature 35.5° C. or 37.5° C., and
wherein the process results in total galactosylation of the omalizumab of 8%±0.8% to 12%±1.2% compared to cell culture process performed at temperature 35.5° C. or 37.5° C. or wherein the process results in expression of acidic variants of the omalizumab of 12%±1.2% to 16%±1.6% compared to cell culture process performed at temperature 35.5° C. or 37.5° C.
11 . The process as claimed in claim 10 , wherein the process is performed in a fed batch mode, or continuous mode or perfusion mode.
12 . The process as claimed in claim 1 , wherein the process is performed in a fed batch mode, or continuous mode or perfusion mode.