Process for manipulating the level of glycan content of a glycoprotein

1. A method for manipulating the fucosylated glycan content on a recombinant protein comprising

inoculating a bioreactor with mammalian host cells expressing the recombinant protein,

culturing the host cells in a serum free, chemically defined cell culture medium; wherein the cell culture medium includes from 10 to 100 ppb copper and from 50 to 1000 nM manganese,

harvesting the recombinant protein produced by the host cell.

2. The method of claim 1 , wherein there is also an increase in the level of β-galactosylation on the recombinant protein.

3. The method according to claim 1 , wherein the concentration of copper is 100 ppb.

4. The method according to claim 1 , wherein the concentration of manganese is 1000 nM.

5. The method according to claim 1 , wherein the fucosylated glycan content is manipulated to influence the level of afucosylated and β-galactosylated glycans without impacting cell culture performance.

6. The method according to claim 1 , further comprising a temperature shift.

7. The method according to claim 6 , wherein the temperature shift is from 36° C. to 31° C.

8. The method according to claim 6 , wherein the temperature shift occurs at the transition between the growth phase and production phase.

9. The method according to claim 6 , wherein the temperature shift occurs during the production phase.

10. The method according claim 1 wherein the host cell expressing the recombinant protein is cultured in a batch culture, fed-batch culture, perfusion culture, or combinations thereof.

11. The method according to claim 10 , wherein the culture is a perfusion culture.

12. The method according to claim 11 , wherein perfusion comprises continuous perfusion.

13. The method according to claim 11 , wherein the rate of perfusion is constant.

14. The method according to claim 11 , wherein the perfusion is performed at a rate of less than or equal to 1.0 working volumes per day.

15. The method according to claim 11 , wherein the perfusion is accomplished by alternating tangential flow.

16. The method according to claim 1 , wherein the bioreactor has a capacity of at least 500 L.

17. The method according to claim 1 wherein the bioreactor has a capacity of at least 500 L to 2000 L.

18. The method according to claim 1 wherein the bioreactor has a capacity of at least 1000 L to 2000 L.

19. The method according to claim 1 , wherein the bioreactor is inoculated with at least 0.5×10 6 cells/mL.

20. The method according to claim 1 , wherein the serum-free chemically defined cell culture medium is a perfusion cell culture medium.

21. The method according to claim 1 , wherein the host cells are Chinese Hamster Ovary (CHO) cells.

22. The method according to claim 1 , wherein the recombinant protein is a glycoprotein.

23. The method according to claim 1 , wherein the recombinant protein is selected from the group consisting of a human antibody, a humanized antibody, a chimeric antibody, a recombinant fusion protein, or a cytokine.

24. The method according to claim 1 , wherein the recombinant protein produced by the host cell is purified and formulated into a pharmaceutically acceptable formulation.