Claims for Patent: 9,090,688

Summary for Patent: 9,090,688

Title:	Methods for controlling the galactosylation profile of recombinantly-expressed proteins
Abstract:	The present invention relates to methods for modulating the glycosylation profile of recombinantly-expressed proteins. In particular, the present invention relates to methods of controlling the galactosylation profile of recombinantly-expressed proteins by supplementing production medium, e.g., a hydrolysate-based or a chemically defined medium, with manganese and/or D-galactose.
Inventor(s):	Bengea; Cornelia (Auburn, MA), Rives; Lisa M. (Natick, MA), Hossler; Patrick (Westborough, MA)
Assignee:	AbbVie Inc. (North Chicago, IL)
Application Number:	14/493,068
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 9,090,688
Patent Claims:	1. A process for producing an antibody comprising the heavy and light chain variable domains of adalimumab, the process comprising culturing a mammalian cell which produces said antibody in a cell culture media that comprises a concentration of manganese of 0.2-100 .mu.M and a concentration of galactose of 1-100 mM, wherein said concentration of manganese and galactose is sufficient to increase the galactosylation level of said antibody as compared to the galactosylation level of an antibody comprising the heavy and light chain variable domains of adalimumab produced in a cell culture media that does not comprise said concentration of manganese and galactose, thereby producing said antibody. 2. The process of claim 1, wherein said culturing is done in a suspension culture. 3. The process of claim 1, wherein the mammalian cell is a CHO cell. 4. The process of claim 1, wherein the mammalian cell is cultured for at least 4 days. 5. The process of claim 1, wherein the process is a fed batch process. 6. The process of claim 1, wherein the media is selected from the group consisting of production media and feed media. 7. The process of claim 1, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 40-100 .mu.M. 8. The process of claim 1, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 10-100 mM. 9. The process of claim 1, wherein the media comprises a sufficient amount of manganese and galactose to achieve a manganese (Mn) and a galactose (Gal) concentration selected from the group consisting of 0.2/1, 0.2/4, 0.2/30, 0.5/1, 0.5/4, 0.5/30, 10/10, 10/20, 10/40, 20/10, 20/20, 20/40, 25/15, 40/10, 40/20, 40/40, 40/100, 50/30, 60/20, 60/40, 60/100, 80/20, 80/40, 80/100, 100/20, 100/40, and 100/100 Mn (.mu.M)/Gal (mM). 10. The process of claim 1, wherein manganese and galactose are each present in the media at a concentration sufficient to produce a composition comprising said antibody in which at least 10% of the total N-linked oligosaccharides present on said antibody are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F). 11. The process of claim 10, wherein at least 15% of the total N-linked oligosaccharides present on said antibody are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F). 12. The process of claim 1, wherein the antibody is adalimumab. 13. The process of claim 1, further comprising recovering said antibody from the cell culture media. 14. The process of claim 13, further comprising purifying said antibody from the cell culture media. 15. The process of claim 14, further comprising quantifying the levels of galactose-containing fucosylated biantennary oligosaccharides (NA1F and NA2F) and/or agalactosyl fucosylated biantennary oligosaccharides (NGA2F and NGA2F-GlcNAc) present on said antibody. 16. The process of claim 1, wherein said culturing is done in a bioreactor. 17. The process of claim 1, wherein the process is a fed batch process. 18. The process of claim 1, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 0.2-40 .mu.M. 19. The process of claim 1, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 40-60 .mu.M. 20. The process of claim 1, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 50-80 .mu.M. 21. The process of claim 1, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 75-100 .mu.M. 22. The process of claim 1, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 1-60 mM. 23. The process of claim 1, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 60-100 mM. 24. The process of claim 1, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 10-60 mM. 25. A process for producing adalimumab, comprising culturing a mammalian cell which produces adalimumab in a cell culture media that comprises a concentration of manganese of 0.2-100 .mu.M and a concentration of galactose of 1-100 mM, wherein said concentration of manganese and galactose is sufficient to increase the galactosylation level of adalimumab as compared to the galactosylation level of adalimumab produced in a cell culture media that does not comprise said concentration of manganese and galactose, thereby producing adalimumab. 26. The process of claim 25, wherein said culturing is done in a suspension culture. 27. The process of claim 25, wherein the media comprises a hydrolysate. 28. The process of claim 25, wherein the mammalian cell is a CHO cell. 29. The process of claim 25, wherein the mammalian cell is cultured for at least 4 days. 30. The process of claim 25, wherein the process is a fed-batch process. 31. The process of claim 25, wherein the media is selected from the group consisting of production media and feed media. 32. The process of claim 25, wherein the media comprises 40-100 .mu.M of manganese. 33. The process of claim 25, wherein the media comprises 10-100 mM of galactose. 34. The process of claim 25, wherein the media comprises a sufficient amount of manganese and galactose to achieve a manganese (Mn) and a galactose (Gal) concentration selected from the group consisting of 0.2/1, 0.2/4, 0.2/30, 0.5/1, 0.5/4, 0.5/30, 10/10, 10/20, 10/40, 20/10, 20/20, 20/40, 25/15, 40/10, 40/20, 40/40, 40/100, 50/30, 60/20, 60/40, 60/100, 80/20, 80/40, 80/100, 100/20, 100/40, and 100/100 Mn (.mu.M)/Gal (mM). 35. The process of claim 34, wherein at least 15% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F). 36. The process of claim 25, wherein manganese and galactose are each present in the media at a concentration sufficient to produce a composition comprising adalimumab in which at least 10% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F). 37. The process of claim 25, wherein said culturing is done in a bioreactor. 38. The process of claim 25, further comprising adding to the media glucose to achieve a glucose concentration of 3 mg/mL or greater. 39. The process of claim 25, further comprising recovering said adalimumab from the cell culture media. 40. The process of claim 39, further comprising purifying said adalimumab from the cell culture media. 41. The process of claim 40, further comprising quantifying the levels of galactose-containing fucosylated biantennary oligosaccharides (NA1F and NA2F) and/or agalactosyl fucosylated biantennary oligosaccharides (NGA2F and NGA2F-GlcNAc) present on said adalimumab. 42. The process of claim 25, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 0.2-40 .mu.M. 43. The process of claim 25, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 40-60 .mu.M. 44. The process of claim 25, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 50-80 .mu.M. 45. The process of claim 25, wherein the media comprises a sufficient amount of manganese to achieve a manganese concentration in the media of 75-100 .mu.M. 46. The process of claim 25, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 1-60 mM. 47. The process of claim 25, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 60-100 mM. 48. The process of claim 25, wherein the media comprises a sufficient amount of galactose to achieve a galactose concentration in the media of 10-60 mM.

Details for Patent 9,090,688

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2031-04-27
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2031-04-27
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2031-04-27
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	09/23/2014		2031-04-27