Use of low temperature and/or low pH in cell culture
View Patent ↗The present disclosure provides a method of reducing protein misfolding and aggregation in the cell culture by growing the cell culture at a reduced temperature and/or reduced pH. As a result, the quality of the protein produced in the cell culture is greatly improved. Thus, the present disclosure facilitates improvements in the efficacy of therapeutic proteins produced in cell culture.
1. A method of improving protein production in a cell culture comprising:
(a) growing cells in the cell culture at a reduced temperature, wherein the reduced temperature is in a range of 27.0° C. to less than 30.0° C.; and
(b) growing cells in the cell culture at a reduced pH, wherein the reduced pH is in a range of 6.80 to less than 7.00;
wherein the cell culture is a mammalian cell culture;
wherein the method reduces production of misfolded proteins and/or aggregated proteins; and
wherein the protein produced is TNFR-Fc or sIL-13R.
2. The method of claim 1 , wherein the cell culture is a large-scale cell culture.
3. The method of claim 1 , wherein the cell culture is a fed-batch cell culture.
4. The method of claim 1 , wherein the cell culture is a CHO cell culture.
5. The method of claim 1 wherein the reduced temperature is 29.5° C.
6. The method of claim 1 , wherein the reduced pH is 6.95.
7. The method of claim 1 , wherein the method modifies the level of protein glycosylation of the produced protein.
8. The method of claim 7 , wherein the protein glycosylation is N-glycan sialylation.
9. The method of claim 1 , further comprising a step of isolating the protein from the cell culture.
10. The method of claim 9 , wherein the protein is further purified or processed for formulation.
11. The method of claim 10 , wherein the protein is formulated into a pharmaceutical composition.