Medium for the protein-free and serum-free cultivation of cells
A medium is described for the protein-free and serum-free cultivation of cells, especially mammalian cells, whereby the medium contains a proportion of soy hydrolysate.
1. An animal protein-free and serum-free mammalian cell culture medium, said medium comprising more than 10 wt % ultrafiltered soy hydrolysate based on the total dry weight of said medium, wherein at least 40% of said soy hydrolysate has a molecular weight of <500 daltons.
2. A medium in accordance with claim 1 , wherein has an endotoxin content of <500 U/g.
3. A medium in accordance with claim 1 , wherein at least 50% of the soy hydrolysate has a molecular weight of ≦500 daltons.
4. A medium in accordance with claim 3 , wherein at least 55% of the soy hydrolysate has a molecular weight of ≦500 daltons.
5. A medium in accordance with claim 1 , wherein said medium also contains an amino acid.
6. A medium in accordance with claim 5 , wherein said amino acid is selected from the group consisting of L-asparagine, L-cysteine, L-cystine, L-proline, L-tryptophan, L-glutamine and mixtures thereof.
7. A medium in accordance with claim 1 wherein said medium also contains auxiliary substances selected from the group consisting of buffer substances, oxidation stabilizers, stabilizers to counteract mechanical stress, and protease inhibitors.
8. A process for the cultivation of mammalian cells, comprising introducing said cells into a medium in accordance with claim 1 and growing said cells in said medium.
9. The process of claim 8 , further comprising introducing mammalian cells selected from the group consisting of CHO cells and BHK cells.
10. A process for the production of a protein from cell culture comprising:
introducing mammalian cells into a medium in accordance with claim 1 , wherein said cells express a desired protein;
growing said cells in said medium and expressing said protein, thereby producing a mixture of said cells and said protein in the medium; and
purifying said protein from said mixture.
11. The process of claim 10 , further comprising introducing cells which produce a recombinant protein selected from the group consisting of Factor II, Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Protein S, Protein C, activated forms of these factors, and vWF.
12. A cell culture composition comprising mammalian cells and a medium in accordance with claim 1 .
13. A process for producing a cell culture medium according to claim 1 , comprising:
obtaining a soy hydrolysate;
ultrafiltering said soy hydrolysate using an ultrafiltration process;
purifying said soy hydrolysate fraction using size exclusion chromatography; and
selecting the soy hydrolysate fractions consisting of soy hydrolysate having a molecular weight ≦500 daltons; and
adding the soy hydrolysate to an animal protein-free and serum-free medium in an amount of more than 10 wt % based on the total dry weight of said medium to produce the cell culture medium.