Cell culture process
Polypeptides having target levels of C-terminal variants are described.
1. A method of producing a preparation of a recombinant antibody, comprising:
culturing a cell in a medium under conditions in which the cell expresses a recombinant antibody, wherein the medium comprises 1.5 g/L lysine to less than 20 g/L lysine; and
isolating the recombinant antibody, thereby producing a preparation of the recombinant antibody.
2. The method of claim 1 , wherein the medium comprises 2 g/L lysine to 19.5 g/L lysine.
3. The method of claim 1 , wherein the medium comprises 5 g/L lysine to 15 g/L lysine.
4. The method of claim 1 , wherein the medium comprises 8 g/L lysine to 12 g/L lysine.
5. The method of claim 1 , wherein the medium comprises 10 g/L lysine.
6. The method of claim 1 , wherein the culturing produces C-terminal variants of the recombinant antibody that differ in amino acid sequence only by the presence or absence of a lysine at their carboxyl termini, and the method further comprises measuring a level of one or more C-terminal variants of the recombinant antibody in the preparation.
7. The method of claim 6 , wherein the C-terminal variants of the recombinant antibody comprise one or more of a K1 lysine variant of the recombinant antibody and a K2 lysine variant of the recombinant antibody.
8. The method of claim 7 , wherein the level of one or more of K1 lysine variant and K2 lysine variant in the preparation is increased relative to a preparation of the recombinant antibody produced using a medium not comprising 1.5 g/L lysine to less than 20 g/L lysine.
9. The method of claim 7 , wherein the preparation comprises a target value of one or more of K1 lysine variants of the recombinant antibody and K2 lysine variants of the recombinant antibody.
10. The method of claim 9 , wherein the target value of K1 lysine variants of the recombinant antibody is at least 10% of the recombinant antibody in the preparation.
11. The method of claim 9 , wherein the target value of K2 lysine variants of the recombinant antibody is at least 4% of the recombinant antibody in the preparation.
12. The method of claim 1 , wherein the host cell is a CHO cell.
13. The method of claim 1 , wherein the antibody is adalimumab.
14. The method of claim 1 , wherein the medium has a pH of about 6.7 to about 7.1.
15. The method of claim 1 , wherein the host cell is cultured at a temperature of about 31° C. to about 37° C.
16. A method of producing a preparation of a recombinant antibody, comprising:
culturing a cell in a medium under conditions in which the cell expresses a recombinant antibody, wherein the medium comprises 1.5 g/L arginine to less than 20 g/L arginine, and wherein the culturing produces C-terminal variants of the recombinant antibody that differ in amino acid sequence only by the presence or absence of a lysine at their carboxyl termini;
isolating the recombinant antibody, thereby producing a preparation of the recombinant antibody; and
measuring a level of one or more C-terminal variants of the recombinant antibody in the preparation.
17. The method of claim 16 , wherein the medium comprises 2 g/L arginine to 19.5 g/L arginine.
18. The method of claim 16 , wherein the medium comprises 5 g/L arginine to 15 g/L arginine.
19. The method of claim 16 , wherein the medium comprises 8 g/L arginine to 12 g/L arginine.
20. The method of claim 16 , wherein the medium comprises 10 g/L arginine.
21. The method of claim 16 , wherein the C-terminal variants of the recombinant antibody comprise one or more of a K1 lysine variant of the recombinant antibody and a K2 lysine variant of the recombinant antibody.
22. The method of claim 21 , wherein the level of one or more of K1 lysine variant and K2 lysine variant in the preparation is increased relative to a preparation of the recombinant antibody produced using a medium not comprising 1.5 g/L to less than 20 g/L arginine.
23. The method of claim 21 , wherein the preparation comprises a target value of one or more of K1 lysine variants of the recombinant antibody and K2 lysine variants of the recombinant antibody.
24. The method of claim 23 , wherein the target value of K1 lysine variants of the recombinant antibody is at least 10% of the recombinant antibody in the preparation.
25. The method of claim 23 , wherein the target value of K2 lysine variants of the recombinant antibody is at least 4% of the recombinant antibody in the preparation.
26. The method of claim 16 , wherein the host cell is a CHO cell.
27. The method of claim 16 , wherein the antibody is adalimumab.
28. The method of claim 16 , wherein the medium has a pH of about 6.7 to about 7.1.
29. The method of claim 16 , wherein the host cell is cultured at a temperature of about 31° C. to about 37° C.
30. A method of producing a preparation of adalimumab, comprising:
culturing a CHO cell in a medium under conditions in which the CHO cell expresses adalimumab, wherein the medium comprises 1.5 g/L lysine to less than 20 g/L lysine, or 1.5 g/L arginine to less than 20 g/L arginine, and wherein the culturing produces C-terminal variants of adalimumab that differ in amino acid sequence only by the presence or absence of a lysine at their carboxyl termini;
isolating the adalimumab, thereby producing a preparation of adalimumab; and
measuring a level of one or more C-terminal variants of adalimumab in the preparation.