Claims for Patent: 10,501,769

Summary for Patent: 10,501,769

Title:	Method for the production of a glycosylated immunoglobulin
Abstract:	Herein is reported a method for the production of an immunoglobulin comprising the following steps: a) providing a eukaryotic cell comprising a nucleic acid encoding the immunoglobulin, b) cultivating the eukaryotic cell in a cultivation medium wherein the amount of glucose available in the cultivation medium per time unit is kept constant and limited to less than 80% of the amount that could maximally be utilized by the cells in the cultivation medium per time unit, and c) recovering the immunoglobulin from the culture.
Inventor(s):	Franze; Reinhard (Penzberg, DE), Hirashima; Chikashi (Tokyo, JP), Link; Thomas (Penzberg, DE), Takagi; Yoshinori (Tokyo, JP), Takuma; Shinya (Tokyo, JP), Tsuda; Yuriko (Tokyo, JP)
Assignee:	Hoffmann-La Roche Inc. (Little Falls, NJ) Chugai Seiyaku Kabushiki Kaisha (Tokyo, JP)
Application Number:	14/844,570
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 10,501,769
Patent Claims:	1. A method for the production of an immunoglobulin comprising a) cultivating a Chinese Hamster Ovary (CHO) cell comprising a nucleic acid encoding the immunoglobulin in a cultivation medium with restricted glucose feeding wherein the amount of glucose available in the cultivation medium per time unit is kept constant in all time units in which restricted glucose feeding is performed and the degree of glucose limitation (DGL) is limited to a single constant value in a range between 0.5 and 0.1, wherein feeding is started once the amount of glucose present in the cultivation medium has dropped to or below a preset value in the cultivation and wherein said cultivation is a fed-batch cultivating, and b) recovering the immunoglobulin. 2. The method according to claim 1, wherein the restricted glucose feeding is started at day 2 or day 3 of the cultivating. 3. The method according to claim 1, wherein the cultivating is at a pH value from pH 6.5 to 7.5. 4. The method according to claim 3, wherein the cultivating is at a pH value from pH 6.9 to 7.3. 5. The method according to claim 4, wherein the cultivating is at a pH value from pH 6.95 to pH 7.05 or at a pH value from pH 7.15 to pH 7.25. 6. The method according to claim 1, wherein the immunoglobulin is an immunoglobulin of class G or class E. 7. The method according to claim 1, wherein cultivating the CHO cell is performed for six to twenty days. 8. The method according to claim 1, wherein the immunoglobulin is an anti-IL-6R antibody. 9. The method according to claim 1, wherein the DGL is limited to a constant value in a range between 0.45 and 0.2. 10. The method according to claim 1, wherein the DGL is limited to 0.4, or 0.2. 11. The method according to claim 8, wherein the anti-IL-6R antibody comprises tocilizumab. 12. The method according to claim 8, wherein the DGL is limited to a constant value in a range between 0.45 and 0.2. 13. The method according to claim 8, wherein the DGL is limited to 0.4, or 0.2. 14. The method according to claim 11, wherein the DGL is limited to a constant value in a range between 0.45 and 0.2. 15. The method according to claim 11, wherein the DGL is limited to 0.4, or 0.2. 16. The method according to claim 1, wherein the fraction of the immunoglobulin with a mannose-5 glycostructure is 10% or less of the sum comprising the amount of the polypeptide with a mannose-5 glycostructure, the amount of the polypeptide G(0) isoform, the amount of the polypeptide G(1) isoform, and the amount of the polypeptide G(2) isoform after 7 days of cultivation. 17. The method according to claim 1, wherein the fraction of the immunoglobulin with a mannose-5 glycostructure is 6% or less of the sum comprising the amount of the polypeptide with a mannose-5 glycostructure, the amount of the polypeptide G(0) isoform, the amount of the polypeptide G(1) isoform, and the amount of the polypeptide G(2) isoform after 7 days of cultivation. 18. A method for the production of an immunoglobulin comprising a) cultivating a Chinese Hamster Ovary (CHO) cell comprising a nucleic acid encoding the immunoglobulin in a cultivation medium with restricted glucose feeding wherein the amount of glucose available in the cultivation medium per time unit is kept constant in all time units in which restricted glucose feeding is performed and the degree of glucose limitation (DGL) is limited to a single constant value in a range between 0.5 and 0.1, and wherein the restricted glucose feeding is started at day 2 or day 3 of the cultivating and wherein said cultivating is a fed-batch cultivating, and b) recovering the immunoglobulin. 19. The method according to claim 1, wherein the time unit is about 1 minute. 20. The method according to claim 1, wherein the time unit is about 1 hour. 21. The method according to claim 1, wherein the time unit is about 6 hours. 22. The method according to claim 1, wherein the time unit is about 12 hours. 23. The method according to claim 1, wherein the time unit is about 24 hours. 24. The method according to claim 1, wherein density of CHO cells at the beginning of the cultivating is about 10.sup.5 cells per milliliter. 25. The method according to claim 1, wherein density of CHO cells at the beginning of the cultivating is about 8.times.10.sup.5 cells per milliliter. 26. The method according to claim 1, wherein density of CHO cells at the beginning of the cultivating is about 10.times.10.sup.5 cells per milliliter. 27. The method according to claim 1, wherein density of CHO cells at the beginning of the cultivating is about 12.times.10.sup.5 cells per milliliter. 28. The method according to claim 1, wherein density of CHO cells is from about 10.sup.5 cells per milliliter to about 60.times.10.sup.5 cells per milliliter during the cultivation. 29. The method according to claim 18, wherein the time unit is about 1 minute. 30. The method according to claim 18, wherein the time unit is about 1 hour. 31. The method according to claim 18, wherein the time unit is about 6 hours. 32. The method according to claim 18, wherein the time unit is about 12 hours. 33. The method according to claim 18, wherein the time unit is about 24 hours. 34. The method according to claim 18, wherein density of CHO cells at the beginning of the cultivating is about 10.sup.5 cells per milliliter. 35. The method according to claim 18, wherein density of CHO cells at the beginning of the cultivating is about 8.times.10.sup.5 cells per milliliter. 36. The method according to claim 18, wherein density of CHO cells at the beginning of the cultivating is about 10.times.10.sup.5 cells per milliliter. 37. The method according to claim 18, wherein density of CHO cells at the beginning of the cultivating is about 12.times.10.sup.5 cells per milliliter. 38. The method according to claim 18, wherein density of CHO cells is from about 10.sup.5 cells per milliliter to about 60.times.10.sup.5 cells per milliliter during the cultivation. 39. The method according to claim 1, wherein cultivating the CHO cell is performed for six to fifteen days. 40. The method according to claim 1, wherein cultivating the CHO cell is performed for six to eight days. 41. The method according to claim 1, wherein the fraction of the immunoglobulin with a mannose-5 glycostructure is 8% or less of the sum comprising the amount of the polypeptide with a mannose-5 glycostructure, the amount of the polypeptide G(0) isoform, the amount of the polypeptide G(1) isoform, and the amount of the polypeptide G(2) isoform after 7 days of cultivation.

Details for Patent 10,501,769

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	ACTEMRA	tocilizumab	Injection	125276	01/08/2010		2029-10-26
Genentech, Inc.	ACTEMRA	tocilizumab	Injection	125472	10/21/2013		2029-10-26
Genentech, Inc.	ACTEMRA	tocilizumab	Injection	125472	11/19/2018		2029-10-26