Claims for Patent: 9,677,105

Summary for Patent: 9,677,105

Title:	Methods of cell culture
Abstract:	Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using ammonium and/or lysine, are described.
Inventor(s):	Collins; Brian Edward (Arlington, MA), Prentice; Holly (Carlisle, MA), Belongia; Brett (North Andover, MA), Tijani; Rasheed (Haverhill, MA)
Assignee:	Momenta Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	13/829,249
Patent Claims:	1. A method of producing an adalimumab preparation having a target value of one or more of high mannose glycans and fucosylated glycans, the method comprising: (a) providing a cell genetically engineered to express adalimumab; (b) culturing the cell in a culture medium comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine under conditions in which the cell expresses the adalimumab; and (c) harvesting a preparation of the adalimumab produced by the cell that meets-the target value of one or more of high mannose glycans and fucosylated glycans; wherein (i) the target value of high mannose glycans is a level that is at least 10% higher than a level of high mannose glycans in a preparation of adalimumab produced by culturing the cell in the medium not comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine, and (ii) the target value of fucosylated glycans is a level that is at least 10% lower than a level of fucosylated glycans in a preparation of adalimumab produced by culturing the cell in the medium not comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine. 2. The method of claim 1, wherein the target value is a level of one or more of high mannose glycans and fucosylated glycans in a reference preparation of adalimumab. 3. The method of claim 1, wherein the target value is a level of one or more of high mannose glycans and fucosylated glycans in a reference preparation of adalimumab. 4. The method of claim 1, further comprising evaluating a level of one or more of high mannose glycans and fucosylated glycans in the adalimumab preparation. 5. The method of claim 1, wherein the target value is one or more of: (a) 0.1% to 20% high mannose glycans, and (b) 70% to 100% fucosylated glycans. 6. The method of claim 1, wherein the culture medium further comprises one or more of glucosamine, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, a peptone, and DMSO. 7. A method of producing an adalimumab preparation, the method comprising: (a) providing a target value of one or more of high mannose glycans and fucosylated glycans; (b) providing a cell genetically engineered to express adalimumab; (c) culturing the cell in a culture medium comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine under conditions in which the cell expresses the adalimumab; (d) harvesting a preparation of the adalimumab produced by the cell; and (e) formulating the preparation into a drug product if the preparation meets the target value of the one or more of high mannose glycans and fucosylated glycans; wherein (i) the target value of high mannose glycans is a level that is at least 10% higher than a level of high mannose glycans in a preparation of adalimumab produced by culturing the cell in the medium not comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine, and (ii) the target value of fucosylated glycans is a level that is at least 10% lower than a level of fucosylated glycans in a preparation of adalimumab produced by culturing the cell in the medium not comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine. 8. The method of claim 7, further comprising evaluating a level of one or more of high mannose glycans and fucosylated glycans in the adalimumab preparation. 9. The method of claim 7, wherein the culture medium further comprises one or more of glucosamine, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, a peptone, and DMSO. 10. A method of increasing a level of high mannose glycans in an adalimumab preparation, the method comprising: (a) providing a cell genetically engineered to express adalimumab; (b) culturing the cell in a culture medium comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine under conditions in which the cell expresses the adalimumab; and (c) harvesting a preparation of the adalimumab prepared by the cell, wherein the preparation has a level of high mannose glycans that is at least 10% higher than a level of high mannose glycans in a preparation of adalimumab produced by culturing the cell in the medium not comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine. 11. The method of claim 10, further comprising evaluating a level of one or more high mannose glycans. 12. The method of claim 10, wherein the culture medium further comprises one or more of glucosamine, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, a peptone, and DMSO. 13. A method of decreasing a level of fucosylated glycans in an adalimumab preparation, the method comprising: (a) providing a cell genetically engineered to express adalimumab; (b) culturing the cell in a culture medium comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine under conditions in which the cell expresses the adalimumab; and (c) harvesting a preparation of the adalimumab produced by the cell, wherein the preparation has a level of fucosylated glycans that is at least 10% lower than a level of fucosylated glycans in a preparation of adalimumab produced by culturing the cell in the medium not comprising 1 mM to 50 mM ammonium and 1 g/L to 30 g/L lysine. 14. The method of claim 13, further comprising evaluating a level of one or more fucosylated glycans. 15. The method of claim 13, wherein the culture medium further comprises one or more of glucosamine, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, a peptone, and DMSO. 16. The method of any one of claims 1, 7, 10, and 13, wherein the cell is a Chinese Hamster Ovary (CHO) cell. 17. The method of claim 4 or 8, further comprising recording the evaluated level of one or more of high mannose glycans and fucosylated glycans in a batch record for the preparation. 18. The method of claim 11, further comprising recording the evaluated level of one or more high mannose glycans in a batch record for the preparation. 19. The method of claim 14, further comprising recording the evaluated level of one or more fucosylated glycans in a batch record for the preparation. 20. The method of any one of claims 1, 7, 10, and 13, wherein the culturing step comprises adding ammonium and lysine to the culture medium. 21. The method of any one of claims 1, 7, 10, and 13, wherein the culturing step comprises a first stage and a second stage, and wherein the first stage comprises culturing the cell in the culture medium comprising a first level of ammonium and a first level of lysine, and the second stage comprises culturing the cell in the culture medium comprising (i) a second level of ammonium which is increased relative to the first level of ammonium and (ii) a second level of lysine which is increased relative to the first level of lysine. 22. The method of claim 21, wherein the second level of ammonium is 1 mM to 50 mM ammonium and the second level of lysine is 1 g/L to 30 g/L lysine. 23. The method of claim 21, wherein the first stage comprises culturing the cell in the first level of ammonium and the first level of lysine for 1 to 8 days. 24. The method of claim 23, wherein the second stage comprises culturing the cell in the second level of ammonium and the second level of lysine for 1 to 12 days.

Details for Patent 9,677,105

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2039-02-26
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2039-02-26
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2039-02-26
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	09/23/2014		2039-02-26
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	11/23/2015		2039-02-26
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	03/09/2016		2039-02-26