Claims for Patent: 10,144,944

Summary for Patent: 10,144,944

Title:	Methods of cell culture
Abstract:	Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described.
Inventor(s):	Prentice; Holly (Carlisle, MA)
Assignee:	Momenta Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	15/496,368
Patent Claims:	1. A method of producing an adalimumab preparation having a target value of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans, the method comprising: (a) providing a cell genetically engineered to express adalimumab; (b) culturing the cell in a culture medium comprising 0.1 mg/L to 10 mg/L putrescine 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 the one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans, wherein the target value of fucosylated glycans, galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of fucosylated glycans, galactosylated glycans, or sialylated glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine; or wherein the target value of high mannose glycans is a level at least 10% lower than a level of high mannose glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 2. The method of claim 1, wherein the target value is a level of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans in a reference preparation of adalimumab. 3. The method of claim 1, further comprising evaluating a level of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans in the adalimumab preparation. 4. The method of claim 1, wherein the target value is one or more of: (a) 70% to 100% fucosylated glycans; (b) 1% to 95% galactosylated glycans; (c) 0.1% to 20% high mannose glycans; and (d) 0.1% to 90% sialylated glycans. 5. The method of claim 1, wherein the culture medium further comprises one or more of lysine, ammonium, DMSO, copper, glucose, a growth factor, a vitamin, a lipid, and a peptone. 6. A method of producing an adalimumab preparation, the method comprising: (a) providing a target value of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans; (b) providing a cell genetically engineered to express adalimumab; (c) culturing the cell in a culture medium comprising 0.1 mg/L to 10 mg/L putrescine under conditions in which the cell expresses the adalimumab; (d) harvesting a preparation of the adalimumab produced by the cell; and (e) formulating the adalimumab preparation into a drug product if the adalimumab preparation meets the target value of the one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans; wherein the target value of fucosylated glycans, galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of fucosylated glycans, galactosylated glycans, or sialylated glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine; or wherein the target value of high mannose glycans is a level at least 10% lower than a level of high mannose glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 7. The method of claim 6, further comprising evaluating a level of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans in the adalimumab preparation. 8. The method of claim 6, wherein the culture medium further comprises one or more of lysine, ammonium, DMSO, copper, glucose, a growth factor, a vitamin, a lipid, and a peptone. 9. A method of increasing a level of one or more of fucosylated glycans, galactosylated glycans and sialylated 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 0.1 mg/L to 10 mg/L putrescine under conditions in which the cell expresses the adalimumab; and (c) harvesting a preparation of adalimumab produced by the cell, wherein the adalimumab preparation has a level of one or more of fucosylated glycans, galactosylated glycans and sialylated glycans that is at least 10% higher than a level of one or more of fucosylated glycans, galactosylated glycans and sialylated glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 10. The method of claim 9, further comprising evaluating a level of one or more of fucosylated glycans, galactosylated glycans and sialylated glycans. 11. The method of claim 9, wherein the culture medium further comprises one or more of lysine, ammonium, DMSO, copper, glucose, a growth factor, a vitamin, a lipid, and a peptone. 12. A method of decreasing a level of one or more 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 0.1 mg/L to 10 mg/L putrescine under conditions in which the cell expresses the adalimumab; and (c) harvesting a preparation of the adalimumab produced by the cell, wherein the adalimumab preparation has a level of high mannose glycans that is at least 10% lower than a level of high mannose glycans in an adalimumab preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 13. The method of claim 12, further comprising evaluating a level of the one or more high mannose glycans. 14. The method of claim 12, wherein the culture medium further comprises one or more of lysine, ammonium, DMSO, copper, glucose, a growth factor, a vitamin, a lipid, and a peptone. 15. The method of any one of claim 1, 6, 9, or 12, wherein the culturing step comprises a first stage and a second stage, wherein the first stage comprises culturing the cell in the culture medium comprising a first level of putrescine, and the second stage comprises culturing the cell in the culture medium comprising a second level of putrescine which is increased relative to the first level. 16. The method of claim 15, wherein the second level is 0.1 mg/L to 10 mg/L putrescine. 17. The method of claim 15, wherein the first stage comprises culturing the cell in the first level of putrescine for 1 to 8 days. 18. The method of claim 17, wherein the second stage comprises culturing the cell in the second level of putrescine for 1 to 12 days. 19. The method of any one of claim 1, 6, 9, or 12, wherein the cell is a Chinese Hamster Ovary (CHO) cell. 20. The method of claim 3 or 7, further comprising recording the evaluated level of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans in a batch record for the preparation. 21. The method of claim 10, further comprising recording the evaluated level of one or more of fucosylated glycans, galactosylated glycans and sialylated glycans in a batch record for the preparation. 22. The method of claim 13, further comprising recording the evaluated level of one or more high mannose glycans in a batch record for the preparation. 23. The method of any one of claim 1, 6, 9, or 12, wherein the culturing step comprises adding putrescine to the culture medium.

Details for Patent 10,144,944

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2033-03-14
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2033-03-14
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2033-03-14
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	09/23/2014		2033-03-14