Claims for Patent: 9,512,214

Summary for Patent: 9,512,214

Title:	Methods to control protein heterogeneity
Abstract:	The instant invention relates to the field of protein production and in particular to controlled protein heterogeneity compositions and processes for controlling 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:	14/194,305
Patent Claims:	1. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising supplementing during a production stage a cell culture medium used in the recombinant expression of said immunoglobulin with a yeast hydrolysate and/or a plant hydrolysate to achieve a yeast hydrolysate concentration in the medium 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-GlcNac) present on the recombinantly-expressed immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) of the immunoglobulin recombinantly-expressed in cell culture medium 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 oligosaccharides (sum of NA1F and NA2F) present on the recombinantly-expressed immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) of the immunoglobulin recombinantly-expressed in cell culture medium 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 adalimumab. 3. The method of claim 2, wherein the method produces at least 2.0 g/L of adalimumab. 4. The method of claim 1, wherein the yeast hydrolysate is selected from the group consisting of Bacto TC Yeastolate, HyPep Yeast Extract and UF Yeast Hydrolysate. 5. The method of claim 1, wherein the plant hydrolysate 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. 6. The method of claim 1, wherein the plant hydrolysate is selected from the group consisting of BBL Phytone Peptone, HyPep 1510, SE50 MAF-UF, UF Soy Hydrolysate, Wheat Peptone E1, HyPep 4601 and Proyield WGE80M Wheat. 7. The method of claim 1, wherein the cell culture medium is supplemented with the plant hydrolysate to achieve a plant hydrolysate concentration from 7 g/L to 15 g/L. 8. The method of claim 1, wherein the cell culture medium is supplemented with the plant hydrolysate to achieve a plant hydrolysate concentration of 10 g/L to 15 g/L. 9. The method of claim 1, wherein the cell culture medium is supplemented with the yeast hydrolysate and the plant hydrolysate to achieve a yeast hydrolysate to plant hydrolysate ratio of 0.25 to 1.55. 10. The method of claim 1, wherein the recombinantly-expressed immunoglobulin is produced by a CHO cell line. 11. The method of claim 1, wherein the cell culture medium comprises yeast and/or plant hydrolysate prior to supplementing the medium with the yeast hydrolysate and/or plant hydrolysate. 12. The method of claim 1, wherein the cell culture medium is substantially free of yeast and/or plant hydrolysate prior to supplementing the medium with the yeast hydrolysate and/or plant hydrolysate. 13. The method of claim 1, wherein assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin comprises assessing the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the recombinantly-expressed immunoglobulin. 14. The method of claim 1, wherein 64%-89% of the total N-linked oligosaccharides present on the immunoglobulin are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-GlcNac) form. 15. The method of claim 1, wherein 64%-69% of the total N-linked oligosaccharides present on the immunoglobulin are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-GlcNac) form. 16. The method of claim 1, wherein assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin comprises assessing the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) on the recombinantly-expressed immunoglobulin. 17. The method of claim 1, wherein 8%-31% of the total N-linked oligosaccharides present on the immunoglobulin are of a galactose containing fucosylated biantennary oligosaccharide (sum of NA1F and NA2F) form. 18. The method of claim 1, wherein 29%-31% of the total N-linked oligosaccharides present on the immunoglobulin are of a galactose containing fucosylated biantennary oligosaccharide (sum of NA1F and NA2F) form. 19. The method of claim 1, wherein the method is a fed batch process. 20. The method of claim 1, further comprising collecting and isolating the produced immunoglobulin. 21. The method of claim 1, wherein the production stage initiates at an initial viable cell density of approximately 0.5.times.10.sup.6 cells/mL. 22. The method of claim 1, wherein the oligosaccharide distribution of the recombinantly-expressed immunoglobulin is assessed after the recombinantly-expressed immunoglobulin has been harvested from the cell culture medium. 23. The method of claim 1, wherein the recombinantly-expressed immunoglobulin is expressed in a mammalian cell. 24. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising supplementing a cell culture media used in the recombinant expression of said immunoglobulin with a yeast hydrolysate supplement and/or a plant hydrolysate supplement; and assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin, wherein the method produces at least 2.5 g/L of said immunoglobulin, thereby controlling the oligosaccharide distribution of said immunoglobulin, wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the recombinantly-expressed immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) of the immunoglobulin 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 oligosaccharides (sum of NA1F and NA2F) present on the recombinantly-expressed immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligosaccharides (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. 25. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising supplementing a cell culture media used in the recombinant expression of said immunoglobulin with a yeast hydrolysate supplement and/or a plant hydrolysate supplement, wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the recombinantly-expressed immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) of the immunoglobulin 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 immunoglobulin are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-GlcNac) form, thereby controlling the oligosaccharide distribution of said immunoglobulin. 26. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising supplementing a cell culture media used in the recombinant expression of said immunoglobulin with a yeast hydrolysate supplement and/or a plant hydrolysate supplement, wherein the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) present on the recombinantly-expressed immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligosaccharides (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, and wherein 29%-31% of the total N-linked oligosaccharides present on the recombinantly-expressed immunoglobulin are of a galactose containing fucosylated biantennary oligosaccharide (sum of NA1F and NA2F) form, thereby controlling the oligosaccharide distribution of said immunoglobulin. 27. The method of any one of claim 24, 25 or 26, wherein the yeast hydrolysate supplement is selected from the group consisting of Bacto TC Yeastolate, HyPep Yeast Extract, and UF Yeast Hydrolysate. 28. The method of any one of claim 24, 25 or 26, 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. 29. The method of any one of claim 24, 25 or 26, 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. 30. The method of claim 29, 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. 31. The method of claim 30, 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. 32. The method of any one of claim 24, 25 or 26, 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. 33. The method of any one of claim 24, 25 or 26, wherein the method is a fed batch process. 34. The method of any one of claim 24, 25 or 26, further comprising collecting and isolating the produced immunoglobulin. 35. The method of claim 24, wherein assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin comprises assessing the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the recombinantly-expressed immunoglobulin. 36. The method of claim 24, wherein 64%-89% of the total N-linked oligosaccharides present on the immunoglobulin are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-GlcNac) form. 37. The method of claim 24, wherein 64%-69% of the total N-linked oligosaccharides present on the immunoglobulin are of an agalactosyl fucosylated biantennary oligosaccharide (sum of NGA2F and NGA2F-GlcNac) form. 38. The method of claim 24, wherein assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin comprises assessing the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) on the recombinantly-expressed immunoglobulin. 39. The method of claim 24, wherein 8%-31% of the total N-linked oligosaccharides present on the immunoglobulin are of a galactose containing fucosylated biantennary oligosaccharide (sum of NA1F and NA2F) form. 40. The method of claim 24, wherein 29%-31% of the total N-linked oligosaccharides present on the immunoglobulin are of a galactose containing fucosylated biantennary oligosaccharide (sum of NA1F and NA2F) form. 41. The method of any one of claim 24, 25 or 26, wherein the cell culture medium comprises yeast and/or plant hydrolysate prior to supplementing the medium with the yeast hydrolysate and/or plant hydrolysate. 42. The method of any one of claim 24, 25 or 26, wherein the cell culture medium is substantially free of yeast and/or plant hydrolysate prior to supplementing the medium with the yeast hydrolysate and/or plant hydrolysate. 43. The method of any one of claim 24, 25 or 26, wherein the recombinantly-expressed immunoglobulin is expressed in a mammalian cell. 44. The method of claim 43, wherein the mammalian cell is a CHO cell. 45. The method of any one of claim 24, 25 or 26, wherein the oligosaccharide distribution of the recombinantly-expressed immunoglobulin is assessed after the recombinantly-expressed immunoglobulin has been harvested from the cell culture media. 46. The method of any one of claim 24, 25 or 26, wherein the immunoglobulin is adalimumab. 47. The method of claim 25 or 26, wherein the method produces at least 2.0 g/L of immunoglobulin.

Details for Patent 9,512,214

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