ADAMTS13 protein cell culture supernatant
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 cell culture supernatant, the supernatant comprising recombinant human ADAMTS13 protein,
wherein the supernatant comprises a specific activity of at least 600 units of ADAMTS13 activity per mg of recombinant human ADAMTS13 protein, as measured using a FRETS-VWF73 assay, and wherein the supernatant comprises at least 3 μM zinc.
2. The cell culture supernatant of claim 1 , wherein the supernatant comprises a specific activity of at least 700 units of ADAMTS13 activity per mg of recombinant human ADAMTS13 protein, as measured using a FRETS-VWF73 assay.
3. The cell culture supernatant of claim 1 , wherein the supernatant comprises a specific activity of at least 800 units of ADAMTS13 activity per mg of recombinant human ADAMTS13 protein, as measured using a FRETS-VWF73 assay.
4. The cell culture supernatant of claim 1 , wherein the supernatant comprises a specific activity of at least 900 units of ADAMTS13 activity per mg of recombinant human ADAMTS13 protein, as measured using a FRETS-VWF73 assay.
5. The cell culture supernatant of claim 1 , wherein the supernatant comprises a specific activity of at least 1000 units of ADAMTS13 activity per mg of recombinant human ADAMTS13 protein, as measured using a FRETS-VWF73 assay.
6. The cell culture supernatant according to claim 1 , wherein the supernatant comprises from 3 μM zinc to 12 μM zinc.
7. The cell culture supernatant according to claim 6 , wherein the supernatant comprises 5 μM zinc.
8. The cell culture supernatant according to claim 1 , wherein the supernatant further comprises from 0.5 mM calcium to 1.5 mM calcium.
9. The cell culture supernatant according to claim 6 , wherein the supernatant further comprises from 0.5 mM calcium to 1.5 mM calcium.
10. The cell culture supernatant according to claim 7 , wherein the supernatant further comprises from 0.5 mM calcium to 1.5 mM calcium.
11. The cell culture supernatant according to claim 1 , wherein the supernatant further comprises from 2 mg/L nicotinamide to 10 mg/L nicotinamide.
12. The cell culture supernatant according to claim 6 , wherein the supernatant further comprises from 2 mg/L nicotinamide to 10 mg/L nicotinamide.
13. The cell culture supernatant according to claim 7 , wherein the supernatant comprises from 2 mg/L nicotinamide to 10 mg/L nicotinamide.
14. The cell culture supernatant according to claim 1 , wherein the supernatant is a mammalian cell culture supernatant.
15. The cell culture supernatant according claim 1 , wherein the supernatant is a CHO cell, BHK cell, or HEK cell culture supernatant.
16. The cell culture supernatant according claim 6 , wherein the supernatant is a CHO cell, BHK cell, or HEK cell culture supernatant.
17. The cell culture supernatant according claim 7 , wherein the supernatant is a CHO cell, BHK cell, or HEK cell culture supernatant.
18. The cell culture supernatant according to claim 1 , wherein the supernatant is a continuous cell culture supernatant.
19. The cell culture supernatant according to claim 18 , wherein the continuous cell culture is chemostatic cell culture.
20. The cell culture supernatant according to claim 6 , wherein the supernatant is a chemostatic cell culture supernatant.
21. The cell culture supernatant according to claim 7 , wherein the supernatant is a chemostatic cell culture supernatant.
22. The cell culture supernatant according to claim 17 , wherein the supernatant is a chemostatic cell culture supernatant.