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AssigneesBaxter Healthcare S.A., Baxter International Inc.
InventorsLeopold Grillberger, Manfred Reiter, Daniel Fleischanderl
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Described herein is a chemostat-like continuous cell culture system that combines certain advantages of perfusion open systems and chemostat open systems to improve the culturing of mammalian cells, e.g., genetically modified cells, particularly in serum-free or chemically-defined media. The continuous culture system described herein involves culturing mammalian cells in a continuous cell culture system, which comprises a cell retention device, wherein the cell culture system has a dilution rate (D) of less than about 2 d−1, and a cell density of less than about 2×107 cell/mL. Also described herein is a method for producing a polypeptide and/or virus of interest in a continuous cell culture, the method comprising culturing mammalian cells expressing the polypeptide and/or virus of interest in a continuous cell culture system, which comprises a cell retention device, wherein the cell culture system has a dilution rate (D) of less than about 2 d−1, and a cell density of less than about 2×107 cell/mL; and recovering the polypeptide and/or virus of interest from medium of the cell culture system.
A method for producing a polypeptide and/or virus of interest in a continuous cell culture, said method comprising
The method of any of the above claims, wherein said dilution rate is between 0.1 and 1.0 d−1.
The method of any of the above claims, wherein said cell density is less than 1×107 cell/mL.
The method of any of the above claims, wherein said cell culture system has a ratio of the dilution rate and specific growth rate (D/μ) between 1.2 and 5.
The method of any of the above claims, wherein said cell culture system has a specific growth rate of between 0.2 d−1 and 0.8 d−1.
The method of any of the above claims, wherein said cells are cultured in said cell culture system for more than 20 days.
The method of any of the above claims, wherein said cells are cultured in said cell culture system for more than 40 days.
The method of any of the above claims, wherein said cells are cultured in said cell culture system for more than 50 days.
The method of any of the above claims, wherein said dilution rate and said cell density are maintained for at least 80% of the time the cells are cultured in said cell culture system.
The method of any of the above claims, wherein said cell retention device comprises a macroporous microcarrier.
The method of any of the above claims, wherein said cells are cultured in a serum-free medium.
The method of any of the above claims, wherein said cells are cultured in at least 250 L of medium.
The method of any of the above claims, wherein said cells are non-anchorage dependent cells.
The method of any of the above claims, further comprising, before the culturing step, pre-culturing the cells in suspension.
The method of any of the above claims, wherein said cells are genetically modified to express said polypeptide and/or virus of interest.
A composition comprising a polypeptide and/or virus of interest produced according to the method of any of the above claims.
A composition comprising a recombinant ADAMTS13 protein produced according to the method of any of the above claims.
Dependencies are inferred from the claim text.