Cell culture medium for ADAMTS protein expression
View Patent ↗The present invention provides culture mediums that are useful for the expression of ADAMTS proteins, such as ADAMTS13. Methods for the expression and purification of ADAMTS proteins are also provided. In some embodiments, the mediums and methods of the invention are useful for the expression of ADAMTS proteins having high specific activities. Also provided are ADAMTS, e.g., ADAMTS13, protein compositions with high specific activities, which are expressed and purified according to the methods provided herein.
1. A method for expressing a disintegrin and metalloproteinase with thrombospondin motifs 13 (ADAMTS13) protein, the method comprising culturing a cell harboring a nucleic acid encoding an ADAMTS13 protein in a culture medium comprising from 3 μM to 12 μM zinc and at least 2 mg/L nicotinamide,
wherein the ADAMTS13 protein expressed according to the method has a greater specific activity as compared to an ADAMTS13 protein expressed in culture medium comprising 1.5 μM zinc.
2. The method of claim 1 , wherein the culture medium comprises at least 4 mg/L nicotinamide.
3. The method of claim 1 , wherein the culture medium comprises at least 6 mg/L nicotinamide.
4. The method of claim 1 , wherein the culture medium comprises from 2 mg/L to 10 mg/L nicotinamide.
5. The method of claim 1 , wherein the culture medium comprises from 4 mg/L to 10 mg/L nicotinamide.
6. The method of claim 1 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.
7. The method of claim 2 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.
8. The method of claim 3 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.
9. The method of claim 4 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.
10. The method of claim 5 , wherein the culture medium further comprises from 0.5 mM to 1.5 mM calcium.
11. The method of claim 1 , wherein the culture medium further comprises at least 1.0 mM calcium.
12. The method of claim 2 , wherein the culture medium further comprises at least 1.0 mM calcium.
13. The method of claim 3 , wherein the culture medium further comprises at least 1.0mM calcium.
14. The method of claim 4 , wherein the culture medium further comprises at least 1.0 mM calcium.
15. The method of claim 5 , wherein the culture medium further comprises at least 1.0 mM calcium.
16. The method of claim 1 , wherein the cell is a mammalian cell line selected from the group consisting of a human cell line, a hamster cell line, and a murine cell line.
17. The method of claim 4 , wherein the cell is a mammalian cell line selected from the group consisting of a human cell line, a hamster cell line, and a murine cell line.
18. The method of claim 5 , wherein the cell is a mammalian cell line selected from the group consisting of a human cell line, a hamster cell line, and a murine cell line.
19. The method of claim 9 , wherein the cell is a mammalian cell line selected from the group consisting of a human cell line, a hamster cell line, and a murine cell line.
20. The method of claim 10 , wherein the cell is a mammalian cell line selected from the group consisting of a human cell line, a hamster cell line, and a murine cell line.
21. The method of claim 1 , wherein the ADAMTS13 protein is a human ADAMTS13 protein.
22. The method of claim 4 , wherein the ADAMTS13 protein is a human ADAMTS13 protein.
23. The method of claim 16 , wherein the ADAMTS13 protein is a human ADAMTS13 protein.
24. The method of claim 17 , wherein the ADAMTS13 protein is a human ADAMTS13 protein.
25. The method of claim 19 , wherein the ADAMTS13 protein is a human ADAMTS13 protein.