Oligopeptide-free cell culture media
The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg/L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg/L of a polyamine.
1. A method for increasing the expression of a recombinant protein in a cell culture, the method comprising culturing mammalian cells that express a recombinant protein in a protein-free medium that does not comprise oligopeptides, wherein the medium comprises DMEM:HAM's F12 (1:1) basal medium, inorganic salts, amino acids, vitamins, ornithine, and putrescine at a concentration of at least 1 mg/L, and wherein the medium is supplemented with Fe(II) and Cu(II) at an amount greater than the amount in basal DMEM: HAM's F12 (1:1).
2. The method according to claim 1 , wherein the medium is supplemented with L-glutamine, ascorbic acid, ethanolamine, sodium selenite, and a non-ionic surfactant.
3. The method according to claim 1 , wherein the medium is supplemented with L-Asparagine, L-Cysteine, L-Cystine, L-Proline, and L-Tryptophan.
4. The method according to claim 1 , wherein the medium comprises putrescine at a concentration of from about 1.0 mg/L to about 20 mg/L.
5. The method according to claim 1 , wherein the medium further comprises one or more from the group consisting of cadaverine, spermidine, spermine, and agmatine.
6. The method according to claim 1 , wherein the medium further comprises spermine.
7. The method according to claim 1 , wherein the medium comprises a buffer substance.
8. The method according to claim 7 , wherein the buffer substance is selected from one or more of sodium bicarbonate, antioxidants, stabilizers, or protease inhibitors.
9. The method according to claim 1 , wherein the recombinant protein is selected from the group of coagulation factor VII, coagulation factor VIII, coagulation factor IX, vWF, ADAMTS13, and furin.
10. The method according to claim 8 , wherein the recombinant protein is coagulation factor VIII.
11. The method according to claim 8 , wherein the recombinant protein is vWF.
12. The method according to claim 1 , wherein the mammalian cells are CHO cells, 293 cells or BHK cells.
13. The method according to claim 1 , wherein the mammalian cell/protein combination is selected from the group consisting of CHO cells/coagulation factor VIII, CHO cells/coagulation factor VII, CHO cells/ADAMTS13, CHO cells/furin, 293 cells/coagulation factor IX.
14. The method according to claim 13 , wherein the mammalian cell/protein combination is CHO cells/coagulation factor VIII.
15. The method according to claim 1 , wherein the putrescine is synthetically produced.
16. The method according to claim 1 , wherein the putrescine originates from a source other than a protein hydrolysate.
17. The method according to claim 1 , wherein the medium is chemically defined.
18. The method according to claim 1 , wherein the mammalian cells are cultured by a method selected from the group of batch-cultivation, feed-batch-cultivation, perfusion-cultivation and chemostat-cultivation.