Claims for Patent: 9,206,390

Summary for Patent: 9,206,390

Title:	Methods to control protein heterogeneity
Abstract:	The instant invention relates to the field of protein production, and in particular to compositions and processes for controlling and limiting the heterogeneity of proteins expressed in host cells.
Inventor(s):	Rives; Lisa M. (Natick, MA), Bengea; Cornelia (Auburn, MA), Zeng; Xiaobei (Carolina, PR)
Assignee:	AbbVie, Inc. (North Chicago, IL)
Application Number:	13/804,220
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 9,206,390
Patent Claims:	1. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising supplementing during a production stage a cell culture media used in the recombinant expression of said immunoglobulin with a yeast hydrolysate supplement and/or a plant hydrolysate supplement to achieve a yeast hydrolysate concentration in the media of at least 11 g/L and/or a plant hydrolysate concentration of at least 7 g/L and assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin, thereby controlling the oligosaccharide distribution of the recombinantly-expressed immunoglobulin, wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) present on the recombinantly-expressed immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) of the immunoglobulin recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement during the production stage; and/or wherein the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) present on the recombinantly-expressed immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) of the immunoglobulin recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement during the production stage. 2. The method of claim 1, wherein the immunoglobulin is an anti-TNF.alpha.antibody. 3. The method of claim 2, wherein the anti-TNF.alpha. antibody is adalimumab. 4. The method of claim 3, wherein the method produces at least 2.0 g/L of adalimumab. 5. The method of claim 1, wherein the yeast hydrolysate supplement is selected from the group consisting of Bacto TC Yeastolate, HyPep Yeast Extract, and UF Yeast Hydrolysate. 6. The method of claim 1, wherein the plant hydrolysate supplement is selected from the group consisting of a soy hydrolysate, a wheat hydrolysate, a rice hydrolysate, a cotton seed hydrolysate, a pea hydrolysate, a corn hydrolysate and a potato hydrolysate. 7. The method of claim 1, wherein the cell culture media is supplemented with a sufficient amount of the plant hydrolysate supplement to achieve a plant hydrolysate concentration of 7 g/L to 15 g/L. 8. The method of claim 7, wherein the cell culture media is supplemented with a sufficient amount of the plant hydrolysate supplement to achieve a plant hydrolysate concentration of 10 g/L to 15 g/L. 9. The method of claim 1, wherein the cell culture media is supplemented with a sufficient amount of the yeast hydrolysate supplement and the plant hydrolysate supplement to achieve a yeast hydrolysate to plant hydrolysate ratio of 0.25 to 1.55. 10. The method of any one of claim 1 or 3, wherein assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin or adalimumab comprises assessing the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) present on the recombinantly-expressed immunoglobulin or adalimumab. 11. The method of any one of claims 1 or 3, wherein 64%-89% of the total N-linked oligosaccharides present on the immunoglobulin or adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-G1cNac) form. 12. The method of any one of claims 1 or 3, wherein 64%-69% of the total N-linked oligosaccharides present on the immunoglobulin or adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-G1cNac) form. 13. The method of any one of claim 1 or 3, wherein assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin or adalimumab comprises assessing the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) on the recombinantly-expressed immunoglobulin or adalimumab. 14. The method of any one of claims 1 or 3, wherein 8%-31% of the total N-linked oligosaccharides present on the immunoglobulin or adalimumab are of a galactose containing fucosylated biantennary oligossacharide (sum of NA1F and NA2F) form. 15. The method of any one of claims 1 or 3, wherein 29%-31% of the total N-linked oligosaccharides present on the immunoglobulin or adalimumab are of a galactose containing fucosylated biantennary oligossacharide (sum of NA1F and NA2F) form. 16. The method of claim 1, wherein the method is a fed batch process. 17. The method of any one of claims 1 or 3, further comprising collecting and isolating the produced immunoglobulin or adalimumab. 18. The method of any one of claims 1 or 3, wherein the production stage initiates at an initial viable cell density of approximately 0.5 .times.10.sup.6 cells/mL. 19. The method of any one of claim 1 or 3, wherein the recombinantly-expressed immunoglobulin or adalimumab is expressed in a mammalian cell. 20. The method of claim 19, wherein the mammalian cell is a CHO cell. 21. The method of any one of claim 1 or 3, wherein the oligosaccharide distribution of the recombinantly-expressed immunoglobulin or adalimumab is assessed after the recombinantly-expressed immunoglobulin or adalimumab has been harvested from the cell culture media. 22. A method for controlling the oligosaccharide distribution of recombinantly-expressed adalimumab comprising supplementing a cell culture media used in the recombinant expression of said adalimumab with a yeast hydrolysate supplement and/or a plant hydrolysate supplement and assessing the oligosaccharide distribution of the recombinantly-expressed adalimumab, wherein the method produces at least 2.5 g/L of adalimumab, thereby controlling the oligosaccharide distribution of said adalimumab, wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) present on the recombinantly-expressed adalimumab is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) of adalimumab recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement; and/or wherein the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) present on the recombinantly-expressed adalimumab is increased as compared to the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) of adalimumab recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement. 23. A method for controlling the oligosaccharide distribution of recombinantly-expressed adalimumab comprising supplementing a cell culture media used in the recombinant expression of said adalimumab with a yeast hydrolysate supplement and/or a plant hydrolysate supplement, wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) present on the recombinantly-expressed adalimumab is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) of adalimumab recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement, and wherein 66%-69% of the total N-linked oligosaccharides present on the recombinantly-expressed adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-G1cNac) form, thereby controlling the oligosaccharide distribution of said adalimumab. 24. A method for controlling the oligosaccharide distribution of recombinantly-expressed adalimumab comprising supplementing a cell culture media used in the recombinant expression of said adalimumab with a yeast hydrolysate supplement and/or a plant hydrolysate supplement, wherein the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) present on the recombinantly-expressed adalimumab is increased as compared to the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) of adalimumab recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement, and wherein 29%-31% of the total N-linked oligosaccharides present on the recombinantly-expressed adalimumab are of a galactose containing fucosylated biantennary oligossacharide (sum of NA1F and NA2F) form, thereby controlling the oligosaccharide distribution of said adalimumab. 25. The method of any one of claim 22, 23 or 24, wherein the yeast hydrolysate supplement is selected from the group consisting of Bacto TC Yeastolate, HyPep Yeast Extract, and UF Yeast Hydrolysate. 26. The method of any one of claim 22, 23 or 24, wherein the plant hydrolysate supplement is selected from the group consisting of a soy hydrolysate, a wheat hydrolysate, a rice hydrolysate, a cotton seed hydrolysate, a pea hydrolysate, a corn hydrolysate and a potato hydrolysate. 27. The method of any one of claim 22, 23 or 24, wherein the cell culture media is supplemented with a sufficient amount of a yeast hydrolysate supplement and/or a plant hydrolysate supplement to achieve a yeast hydrolysate concentration in the media of at least 11 g/L and/or a plant hydrolysate concentration of at least 7 g/L. 28. The method of claim 27, wherein the cell culture media is supplemented with a sufficient amount of the plant hydrolysate supplement to achieve a plant hydrolysate concentration of 7 g/L to 15 g/L. 29. The method of claim 28, wherein the cell culture media is supplemented with a sufficient amount of the plant hydrolysate supplement to achieve a plant hydrolysate concentration of 10 g/L to 15 g/L. 30. The method of any one of claim 22, 23 or 24, wherein the cell culture media is supplemented with a sufficient amount of the yeast hydrolysate supplement and the plant hydrolysate supplement to achieve a yeast hydrolysate to plant hydrolysate ratio of 0.25 to 1.55. 31. The method of any one of claim 22, 23 or 24, wherein the method is a fed batch process. 32. The method of any one of claim 22, 23 or 24, further comprising collecting and isolating the produced adalimumab. 33. The method of claim 22, wherein assessing the oligosaccharide distribution of the recombinantly-expressed adalimumab comprises assessing the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) present on the recombinantly-expressed adalimumab. 34. The method of claim 22, wherein 64%-89% of the total N-linked oligosaccharides present on the adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-G1cNac) form. 35. The method of claim 22, wherein 64%-69% of the total N-linked oligosaccharides present on the adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-G1cNac) form. 36. The method of claim 22, wherein assessing the oligosaccharide distribution of the recombinantly-expressed adalimumab comprises assessing the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) on the recombinantly-expressed adalimumab. 37. The method of claim 22, wherein 8%-31% of the total N-linked oligosaccharides present on the adalimumab are of a galactose containing fucosylated biantennary oligossacharide (sum of NA1F and NA2F) form. 38. The method of claim 22, wherein 29%-31% of the total N-linked oligosaccharides present on the adalimumab are of a galactose containing fucosylated biantennary oligossacharide (sum of NA1F and NA2F) form. 39. The method of any one of claim 22, 23 or 24, wherein the recombinantly-expressed adalimumab is expressed in a mammalian cell. 40. The method of claim 39, wherein the mammalian cell is a CHO cell. 41. The method of any one of claim 22, 23 or 24, wherein the oligosaccharide distribution of the recombinantly-expressed adalimumab is assessed after the recombinantly-expressed adalimumab has been harvested from the cell culture media.

Details for Patent 9,206,390

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2032-09-02
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2032-09-02
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2032-09-02
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	09/23/2014		2032-09-02
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	11/23/2015		2032-09-02
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	03/09/2016		2032-09-02
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	10/17/2016		2032-09-02