Claims for Patent: 8,956,830

Summary for Patent: 8,956,830

Title:	Methods of cell culture
Abstract:	Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using DMSO, are described.
Inventor(s):	Prentice; Holly (Carlisle, MA), Tijani; Rasheed (Haverhill, MA), Belongia; Brett (North Andover, MA)
Assignee:	Momenta Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	13/829,292
Patent Claims:	1. A method of producing a recombinant protein preparation having target values of high mannose glycans and fucosylated glycans, the method comprising: (a) providing a mammalian cell genetically engineered to express a recombinant protein; (b) culturing the cell in a culture medium comprising dimethylsulfoxide (DMSO) 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 values of the high mannose glycans and fucosylated glycans, wherein the target value of high mannose glycans is a level at least 20% higher than a level of high mannose glycans and the target value of fucosylated glycans is a level at least 10% lower than in a preparation produced by culturing the cell in the medium not comprising DMSO. 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 3, wherein the recombinant therapeutic antibody 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. 5. The method of claim 1, further comprising evaluating the level of one or more of high mannose glycans and fucosylated glycans in the recombinant protein preparation. 6. The method of claim 1, wherein the culture medium comprises 0.5% to 5% DMSO. 7. The method of claim 1, wherein the culture medium further comprises one or more of glucosamine, lysine, ammonium, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, and a peptone. 8. A method of producing a recombinant protein preparation, the method comprising: (a) providing target values of mannose glycans and fucosylated glycans; (b) providing a mammalian cell genetically engineered to express a recombinant protein; (c) culturing the cell in a culture medium comprising DMSO 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 values of the high mannose glycans and fucosylated glycans, and wherein the target value of high mannose glycans is a level at least 20% higher than a level of high mannose glycans and the target value of fucosylated glycans is a level at least 10% lower than in a preparation produced by culturing the cell in the medium not comprising DMSO. 9. The method of claim 8, further comprising evaluating the level of the one or more of high mannose glycans and fucosylated glycans in the recombinant protein preparation. 10. The method of claim 8, wherein the culture medium further comprises one or more of glucosamine, lysine, ammonium, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, and a peptone. 11. A method of increasing a level of one or more of sialylated glycans and 3,3,1,0,0 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 DMSO 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 an increased level of one or more of sialylated glycans and 3,3,1,0,0 glycans relative to a level of one or more of sialylated glycans and 3,3,1,0,0 glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising DMSO. 12. The method of claim 11, further comprising evaluating the level of the one or more of sialylated glycans and 3,3,1,0,0 glycans. 13. The method of claim 11, wherein the increased level of the one or more of sialylated glycans and 3,3,1,0,0 glycans is higher than the level of one or more of sialylated glycans and 3,3,1,0,0 glycans in the preparation produced by culturing the cell in the medium not comprising DMSO by at least 10%. 14. The method of claim 11, wherein the culture medium further comprises one or more of glucosamine, lysine, ammonium, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, and a peptone. 15. A method of decreasing a level of one or more fucosylated 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 DMSO 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 decreased level of at least 10% of one or more fucosylated glycans relative to a level of one or more fucosylated glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising DMSO. 16. The method of claim 15, wherein the one or more fucosylated glycans comprise G0F glycans. 17. The method of claim 15, further comprising evaluating the level of one or more of fucosylated glycans. 18. The method of claim 15, wherein the culture medium further comprises one or more of glucosamine, lysine, ammonium, copper, putrescine, glucose, a growth factor, a vitamin, a lipid, and a peptone. 19. The method of any one of claims 1, 8, 11, and 15, wherein the culturing step comprises a first stage and a second stage. 20. The method of claim 19, wherein the first stage comprises culturing the cell in the culture medium comprising a first level of DMSO, and the second stage comprises culturing the cell in the culture medium comprising a second level of DMSO. 21. The method of claim 20, wherein the first level of DMSO is no DMSO and the second level of DMSO is 1% to 5% DMSO. 22. The method of claim 20, wherein the first stage comprises culturing the cell in the first level of DMSO for 1 to 8 days. 23. The method of claim 22, wherein the second stage comprises culturing the cell in the second level of DMSO for 1 to 12 days. 24. The method of any one of claims 1, 8, 11, and 15, wherein the cell is a Chinese Hamster Ovary (CHO) cell. 25. A method of producing a recombinant protein preparation having a target value of high mannose glycans, the method comprising: (a) providing a mammalian cell genetically engineered to express a recombinant protein; (b) culturing the cell in a culture medium comprising DMSO 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 high mannose glycans, wherein the target value is a level at least 20% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 26. The method of claim 25, wherein the recombinant protein is a recombinant therapeutic antibody. 27. The method of claim 25, wherein the target value is a level at least 30% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 28. The method of claim 25, wherein the target value is a level at least 50% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 29. The method of claim 25, wherein the target value is a level at least 70% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 30. The method of claim 25, wherein the target value is a level at least 100% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 31. The method of claim 25, wherein the culture medium comprises 0.5% to 5% DMSO. 32. The method of claim 25, wherein the culture medium comprises 1% to 5% DMSO. 33. The method of claim 25, wherein the culture medium comprises 2% to 5% DMSO. 34. The method of claim 25, wherein the culture medium comprises 3% to 5% DMSO. 35. The method of claim 25, wherein the cell is a CHO cell. 36. The method of claim 25, further comprising evaluating the level of high mannose glycans in the recombinant protein preparation. 37. A method of producing a recombinant protein preparation, the method comprising: (a) providing a mammalian cell genetically engineered to express a recombinant protein; (b) culturing the cell in a culture medium comprising DMSO under conditions in which the cell expresses the recombinant protein; (c) harvesting a preparation of the recombinant protein produced by the cell; and (d) formulating the preparation into a drug product if the preparation comprises a level of high mannose glycans at least 20% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 38. The method of claim 37, wherein the recombinant protein is a recombinant therapeutic antibody. 39. The method of claim 37 or 38, wherein the preparation is formulated into a drug product if the preparation comprises a level of high mannose glycans at least 30% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 40. The method of claim 37 or 38, wherein the preparation is formulated into a drug product if the preparation comprises a level of high mannose glycans at least 50% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 41. The method of claim 37 or 38, wherein the preparation is formulated into a drug product if the preparation comprises a level of high mannose glycans at least 70% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 42. The method of claim 37 or 38, wherein the preparation is formulated into a drug product if the preparation comprises a level of high mannose glycans at least 100% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 43. The method of claim 37 or 38, wherein the culture medium comprises 0.5% to 5% DMSO. 44. The method of claim 37 or 38, wherein the culture medium comprises 1% to 5% DMSO. 45. The method of claim 37 or 38, wherein the culture medium comprises 2% to 5% DMSO. 46. The method of claim 37 or 38, wherein the culture medium comprises 3% to 5% DMSO. 47. The method of claim 37 or 38, wherein the cell is a CHO cell. 48. The method of claim 37 or 38, further comprising evaluating the level of high mannose glycans in the recombinant protein preparation. 49. The method of any one of claim 1, 8, 11, or 15, wherein the culture medium comprises 1% to 5% DMSO. 50. The method of any one of claim 1, 8, 11, or 15, wherein the culture medium comprises 2% to 5% DMSO. 51. The method of any one of claim 1, 8, 11, or 15, wherein the culture medium comprises 3% to 5% DMSO. 52. A method of producing a recombinant therapeutic protein preparation, the method comprising: (a) providing a mammalian cell genetically engineered to express a recombinant therapeutic protein; (b) culturing the cell in a culture medium comprising 1-5% DMSO under conditions in which the cell expresses the recombinant therapeutic protein; (c) harvesting a preparation of the recombinant protein produced by the cell; and (d) formulating the preparation into a therapeutic drug product if the preparation comprises a level of high mannose glycans at least 20% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 53. The method of claim 52, wherein the formulating step comprises formulating the preparation into a therapeutic drug product if the preparation comprises a level of high mannose glycans at least 50% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 54. The method of claim 52, wherein the formulating step comprises formulating the preparation into a therapeutic drug product if the preparation comprises a level of high mannose glycans at least 100% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO. 55. The method of any one of claims 52-54, further comprising evaluating the level of high mannose glycans in the recombinant therapeutic protein preparation. 56. The method of claim 55, further comprising recording the evaluated level of high mannose glycans in a batch record for the preparation. 57. The method of claim 52, wherein the 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.

Details for Patent 8,956,830

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
Idec Pharmaceuticals Corp.	RITUXAN	rituximab	Injection	103737	02/19/2002		2039-02-26