MEDIUM SUPPLEMENTS FOR IMPROVED PROCESS PERFORMANCE
The present invention pertains to a cell culture medium comprising dextran sulfate or a mixture of dextran sulfate and ferric citrate, and methods of using thereof. The present invention further pertains to a method of producing a protein of interest in a large scale cell culture, comprising supplementing the cell culture with dextran sulfate or a mixture of dextran sulfate and ferric citrate.
1 . A large-scale cell culture comprising mammalian cells expressing a polypeptide of interest cells in a cell culture medium, the culture medium comprising between 0.01 g/L and 5 g/L dextran sulfate, and supporting expression by the cells of a polypeptide of interest.
2 . The cell culture of claim 1 , the culture medium further comprising ferric citrate.
3 . The cell culture of claim 2 , the culture medium further comprising 1 mM to 50 mM ferric citrate.
4 . The cell culture of claim 1 , wherein the culture is supplemented with a feed medium between 1 and 25 times.
5 . The cell culture of claim 1 , wherein the lactate production of the cells is lower than the lactate production of cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
6 . The cell culture of claim 1 , wherein the lactate concentration of the culture is between 0.1 g/L and 6 g/L.
7 . The cell culture of claim 1 , wherein the ammonium production of the cells is lower than the ammonium production of cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
8 . The cell culture method of claim 1 , wherein the ammonium concentration of the culture is between 0.1 mM and 20 mM.
9 . The cell culture of claim 1 , wherein the cell-specific lactate production rate to the cell-specific glucose uptake rate ratio (LPR/GUR ratio) of the cells is between 0.5 and 0.5.
10 . The cell culture of claim 1 , wherein the cells are CHO cells, HEK 293 cells, NSO cells, PER.C6 cells, HeLa cells, MDCK cells, or hybridoma cells.
11 . The cell culture of claim 1 , wherein the cells have been adapted to grow in a serum-free medium, a animal protein-free medium, or a chemically defined medium.
12 . The cell culture of claim 1 , wherein the cells have been genetically modified.
13 . The cell culture of claim 1 , wherein the cells produce a polypeptide of interest which is an antibody, a Transforming Growth Factor (TGF) beta superfamily signaling molecule, an Fc fusion protein, a viral protein, or a clotting factor.
14 . The cell culture of claim 1 , wherein the total amount of polypeptide produced by the cells is higher than the total amount of polypeptide produced by cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
15 . The cell culture of claim 1 , wherein the specific productivity of the cells is higher than the specific productivity of cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
16 . The cell culture of claim 1 , wherein the culture is a perfusion culture or a fed batch culture.
17 . The cell culture of claim 1 , wherein the medium is a serum-free medium, AN animal protein-free medium, or a chemically defined medium.
18 . The cell culture of claim 1 , wherein the cells produce a polypeptide of interest which is an antibody, a Transforming Growth Factor (TGF) beta superfamily signaling molecule, an Fc fusion protein, or a clotting factor.
19 . A polypeptide produced in a large-scale cell culture, the culture comprising mammalian cells expressing the polypeptide in a cell culture medium under conditions that support expression of the polypeptide of interest,
wherein the cell culture medium comprises between 0.01 g/L and 5 g/L dextran sulfate, and
wherein the polypeptide is an antibody, a Transforming Growth Factor (TGF) beta superfamily signaling molecule, an Fc fusion protein, a viral protein, or a clotting factor.
20 . The polypeptide of claim 19 , wherein the medium further comprises ferric citrate.
21 . The polypeptide of claim 20 , wherein the culture medium further comprising 1 mM to 50 mM ferric citrate.
22 . The polypeptide of claim 19 , wherein the cells are maintained in the culture for between 1 day and 25 days.
23 . The polypeptide of claim 19 , wherein the cells are maintained longer than cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
24 . The polypeptide of claim 19 , wherein the lactate production of the cells is lower than the lactate production of cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
25 . The polypeptide of claim 19 , wherein the lactate concentration of the culture is between 0.1 g/L and 6 g/L.
26 . The polypeptide of claim 19 , wherein the ammonium production of the cells is lower than the ammonium production of cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
27 . The polypeptide of claim 19 , wherein the ammonium concentration of the culture is between 0.1 mM and 20 mM.
28 . The polypeptide of claim 19 , wherein the cell specific lactate production rate to the cell specific glucose uptake rate ratio (LPR/GUR ratio) of the cells is between 0.5 and 0.5.
29 . The polypeptide of claim 19 , wherein the cells are CHO cells, HEK 293 cells, NSO cells, PER.C6 cells, HeLa cells, MDCK cells, or hybridoma cells.
30 . The polypeptide of claim 19 , wherein the cells have been adapted to grow in serum-free medium, an animal protein-free medium, or a chemically defined medium.
31 . The polypeptide method of claim 19 , wherein the cells have been genetically modified.
32 . The polypeptide of claim 19 , wherein the total amount of polypeptide produced by the cells is higher than the total amount of polypeptide produced by cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
33 . The polypeptide of claim 19 , wherein the specific productivity of the cells is higher than the specific productivity of cells maintained in a culture medium that is substantially free from dextran sulfate and ferric citrate.
34 . The polypeptide of claim 19 , wherein the culture is a perfusion culture or a fed batch culture.
35 . The polypeptide of claim 19 , wherein the medium is a serum-free medium, an animal protein-free medium, or a chemically defined medium.
36 . The polypeptide of claim 19 , which has been isolated from the cell culture.
37 . A large-scale cell culture comprising mammalian cells expressing a virus of interest comprising cells in a cell culture medium, the culture medium comprising between 0.01 g/L and 5 g/L dextran sulfate, and supporting expression by the cells of a polypeptide of interest.