Claims for Patent: 9,217,168

Summary for Patent: 9,217,168

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:	13/829,333
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 9,217,168
Patent Claims:	1. A method of producing a recombinant protein preparation having a target value of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans, the method comprising: (a) providing a cell genetically engineered to express a recombinant protein; (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 recombinant protein; and (c) harvesting a preparation of the recombinant protein produced by the cell that meets the target value of the one or more of galactosylated glycans, high mannose glycans, and sialylated glycans, wherein the target value of galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of galactosylated glycans, or sialylated glycans in a 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 a 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 recombinant protein is a recombinant therapeutic protein. 3. The method of claim 1, wherein the recombinant protein is a recombinant therapeutic antibody. 4. The method of claim 1, wherein the target value is a level of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans in a reference therapeutic protein. 5. The method of claim 1, wherein the target value is a level of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans in a reference therapeutic antibody. 6. The method of claim 4, wherein the reference therapeutic protein is selected from the group consisting of: abatacept, abciximab, adalimumab, aflibercept, alefacept, alemtuzumab, basiliximab, bevacizumab, belatacept, certolizumab, cetuximab, daclizumab, eculizumab, efalizumab, entanercept, gemtuzumab, ibritumomab, infliximab, muromonab-CD3, natalizumab, omalizumab, palivizumab; panitumumab, ranibizumab, rilonacept, rituximab, tositumomab, and trastuzumab. 7. The method of claim 1, further comprising evaluating a level of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans in the recombinant protein preparation. 8. The method of claim 1, wherein the target value is one or more of: (a) 1% to 95% galactosylated glycans; (b) 0.1% to 20% high mannose glycans; and (c) 0.1% to 90% sialylated glycans. 9. 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. 10. A method of producing a recombinant protein preparation, the method comprising: (a) providing a target value of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans; (b) providing a cell genetically engineered to express a recombinant protein; (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 recombinant protein; (d) harvesting a preparation of the recombinant protein 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 galactosylated glycans, high mannose glycans, and sialylated glycans; wherein the target value of galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of galactosylated glycans or sialylated glycans in a 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 a preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 11. The method of claim 10, further comprising evaluating a level of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans in the recombinant protein preparation. 12. The method of claim 10, 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. 13. A method of increasing a level of one or more of galactosylated glycans, and sialylated glycans in a recombinant protein preparation, the method comprising: (a) providing a cell genetically engineered to express a recombinant protein; (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 recombinant protein; and (c) harvesting a preparation of the recombinant protein produced by the cell, wherein the preparation has a level of one or more of galactosylated glycans, and sialylated glycans that is at least 10% higher than a level of one or more of galactosylated glycans and sialylated glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 14. The method of claim 13, further comprising evaluating a level of one or more of galactosylated glycans, and sialylated glycans. 15. The method of claim 13, 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. 16. A method of decreasing a level of one or more high mannose glycans in a recombinant protein preparation, the method comprising: (a) providing a cell genetically engineered to express a recombinant protein; (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 recombinant protein; and (c) harvesting a preparation of the recombinant protein produced by the cell, wherein the preparation has a level of high mannose glycans that is at least 10% lower than a level of high mannose glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine. 17. The method of claim 16, further comprising evaluating a level of the one or more high mannose glycans. 18. The method of claim 16, 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. 19. The method of any one of claim 1, 10, 13, or 16, 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. 20. The method of claim 19, wherein the second level is 0.1 mg/L to 10 mg/L putrescine. 21. The method of claim 19, wherein the first stage comprises culturing the cell in the first level of putrescine for 1 to 8 days. 22. The method of claim 21, wherein the second stage comprises culturing the cell in the second level of putrescine for 1 to 12 days. 23. The method of any one of claim 1, 10, 13, or 16, wherein the cell is a Chinese Hamster Ovary (CHO) cell. 24. The method of claim 7 or 11, further comprising recording the evaluated level of one or more of galactosylated glycans, high mannose glycans, and sialylated glycans in a batch record for the preparation. 25. The method of claim 14, further comprising recording the evaluated level of one or more of galactosylated glycans, and sialylated glycans in a batch record for the preparation. 26. The method of claim 17, further comprising recording the evaluated level of one or more high mannose glycans in a batch record for the preparation. 27. The method of any one of claims 1, 10, 13, and 16, wherein the recombinant protein is selected from the group consisting of: abatacept, abciximab, adalimumab, aflibercept, alefacept, alemtuzumab, basiliximab, bevacizumab, belatacept, certolizumab, cetuximab, daclizumab, eculizumab, efalizumab, entanercept, gemtuzumab, ibritumomab, infliximab, muromonab-CD3, natalizumab, omalizumab, palivizumab, panitumumab, ranibizumab, rilonacept, rituximab, tositumomab, and trastuzumab. 28. The method of any one of claims 1, 10, 13, and 16, wherein the culturing step comprises adding putrescine to the culture medium.

Details for Patent 9,217,168

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Centocor Ortho Biotech Products, L.p.	ORTHOCLONE OKT3	muromanab-cd3	Injection	103463	09/14/1992		2039-02-26
Janssen Biotech, Inc.	REOPRO	abciximab	Injection	103575	12/22/1994		2039-02-26
Genentech, Inc.	RITUXAN	rituximab	Injection	103705	11/26/1997		2039-02-26