Use of depth filtration in series with continuous centrifugation to clarify mammalian cell cultures
View Patent ↗Methods for clarification of cell samples using centrifugation in combination with depth filtration.
1. A method for clarification of an industrial scale cell sample comprising the steps of:
(a) centrifuging a cell sample using a gravitational force within a range of about 8,000×g to about 15,000×g and a Q/Σ ratio ranging between 0.9×10 −8 and 2.8×109 for industrial scale production of therapeutic proteins, whereby a solid phase comprising cells and debris is separated from a centrate; and
(b) applying the centrate to a depth filtration means.
2. The method of claim 1 , wherein the cell sample comprises bacterial cells.
3. The method of claim 1 , wherein the cell sample comprises mammalian cells.
4. The method of claim 3 , wherein the mammalian cells are CHO cells.
5. The method of claim 3 , wherein the mammalian cells are NSO cells.
6. The method of claim 1 , wherein the cell sample comprises a cell suspension, a cell slurry, or a cell culture.
7. The method of claim 1 , wherein the cell sample has a volume of at least about 2200 L.
8. The method of claim 1 , wherein the cell sample has a volume of at least about 15,000 L.
9. The method of claim 1 , wherein the cell sample further comprises an antifoam agent.
10. The method of claim 1 , wherein the centrifuging is performed using a gravitational force within the range of about 8,000×g to about 10,000×g.
11. The method of claim 1 , wherein the separation efficiency is at least about 95% following the centrifuging step.
12. The method of claim 1 , wherein the centrate is substantially free of cells and debris greater than about 2 μm.
13. The method of claim 1 , wherein the depth filtration means comprises one or more filters.
14. The method of claim 13 , wherein the depth filtration means of step (b) is selected from the group consisting of a depth filter and a polishing filter.
15. The method of claim 14 , wherein a final filter of the depth filtration means has a pore size of about 0.1 μm to about 0.2 μm.
16. The method of claim 15 , wherein the final filter has a pore size of at least about 0.2 μm.
17. The method of claim 15 , wherein the final filter has a pore size of at least about 0.1 μm.