Production of growth hormone in serum-free cell culture medium for mammalian cells
The invention relates to a process for the manufacturing of a protein, such as growth hormone, in mammalian cells cultured in a serum-free medium (medium free of components derived from animal serum). The medium contains cobalt and, optionally, cadmium or cyclodextrin.
1. A process for the production of growth hormone comprising the step of culturing cells of a cell line expressing growth hormone in a cell culture medium free of components derived from animal serum, the medium comprising cobalt in a concentration ranging from about 0.5 μM to about 20 μM and cadmium at a concentration of about 10 nM to about 100 nM, wherein the growth hormone is expressed under the control of a metallothionein promoter.
2. The process according to claim 1 , wherein said medium comprises cobalt at a concentration of: a) 1 μM to about 10 μM; b) 5 μM to about 15 μM; c) 7.5 μM to about 12 μM; d) 0.5 μM to about 2.5 μM; e) 0.5 μM to about 2 μM; f) 0.75 μM to about 1.5 μM; g) about 1.0 μM; h) 0.5 μM to about 12.5 μM; i) about 10 μM; or j) about 20 μM.
3. The process according to claim 1 , wherein said medium comprises cadmium at a concentration of about 20 nM to about 100 nM.
4. The process according to claim 1 , wherein said medium comprises cadmium at a concentration of about 50 nM to about 100 nM.
5. The process according to claim 1 , wherein said medium comprises cobalt at a concentration of about 1 μM and cadmium at a concentration of about 100 nM.
6. The process according to claim 1 , wherein said medium further comprises methyl-beta-cyclodextrin at a concentration of about 0.02 mM to about 0.1 mM.
7. The process according to claim 6 , wherein said medium comprises methyl-beta-cyclodextrin at a concentration of about 0.04 mM to about 0.1 mM; or about 0.07 mM to about 0.1 mM.
8. The process according to claim 1 , wherein said medium further comprises ferric ions at a concentration of 5 μM or 6 μM, zinc at a concentration ranging from 0.2 μM to 0.5 μM and copper at a concentration of 25 nM.
9. The process according to claim 8 , wherein the medium comprises zinc at a concentration of 0.2 μM or 0.5 μM.
10. The process according to claim 1 , further comprising collecting the growth hormone from the cell culture.
11. The process according to claim 1 , further comprising purifying the growth hormone.
12. The process according to claim 11 , further comprising formulating the purified growth hormone with a pharmaceutically acceptable carrier to obtain a pharmaceutical composition.
13. The process according to claim 1 , wherein the growth hormone is human growth hormone.
14. A process for the production of growth hormone comprising the step of culturing cells of a cell line expressing growth hormone in a cell culture medium free of components derived from animal serum, the medium comprising cadmium at a concentration of about 10 nM to about 100 nM, wherein the growth hormone is expressed under the control of a metallothionein promoter.
15. The process according to claim 14 , wherein said medium comprises cadmium at a concentration of about 50 nM to about 100 nM.
16. A process for the production of growth hormone comprising the step of culturing cells of a cell line expressing growth hormone in a cell culture medium free of components derived from animal serum, the medium comprising methyl-beta-cyclodextrin at a concentration of about 0.02 mM to about 0.1 mM, wherein the growth hormone is expressed under the control of a metallothionein promoter.
17. The process according to claim 16 , wherein said medium comprises methyl-beta-cyclodextrin at a concentration of: about 0.04 mM to about 0.1 mM; or about 0.07 mM to about 0.1 mM.
18. The process according to claim 1 , wherein the metallothionein promoter is the mouse MT-1 promoter.
19. The process according to claim 1 , wherein the cells are mouse C127 cells.