Claims for Patent: 10,435,464

Summary for Patent: 10,435,464

Title:	Methods for making recombinant proteins
Abstract:	The present invention provides methods and compositions for making proteins, preferably antibodies, more preferably anti-tumor necrosis factor alpha antibodies, and most preferably adalimumab. The present invention further provides methods and compositions for mammalian cell culture, preferably Chinese Hamster Ovary cells.
Inventor(s):	Puchacz; Elzbieta Wiktoria (Pleasanton, CA)
Assignee:
Application Number:	14/845,440
Patent Claims:	1. A method of producing adalimumab in a mammalian cell culture comprising: a) inoculating a fed-batch bioreactor containing a cell culture production medium with mammalian cells comprising a nucleic acid encoding adalimumab or a fragment thereof wherein the mammalian cells are at a concentration from about 0.1 million to about 2 million cells/milliliter, wherein the production medium comprises glucose at an initial concentration from about 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter, such that adalimumab is produced; b) maintaining the glucose in the production medium at a concentration from about 0.1 to about 0.9 grams/liter; c) optionally, maintaining the at least one other hexose in the production medium at a concentration from about 0.1 to about 20 grams/liter; d) optionally, supplementing the production medium with at least one additional feed absent glucose; and e) harvesting the mammalian cell culture at a cell viability from about 20% to about 100%; and f) deactivation, inactivation, or removal of virus from the cell culture. 2. The method of claim 1 wherein the mammalian cells are Chinese Hamster Ovary cells. 3. The method of claim 1 or 2, wherein the adalimumab or the fragment thereof is secreted into the production medium. 4. The method of claim 3 further comprising purifying the adalimumab or the fragment thereof from a production medium. 5. The method of claim 4, further comprising formulating the purified adalimumab or the fragment thereof into a pharmaceutical composition. 6. The method of claim 1, wherein the cell viability is from about 50% to about 80%. 7. A method of producing adalimumab in a mammalian cell culture comprising: a) a growth phase comprising: i) inoculating a perfusion bioreactor, containing a cell culture growth medium, with mammalian cells comprising a nucleic acid encoding adalimumab or a fragment thereof wherein the mammalian cells are at a concentration from about 0.1 million to about 5 million cells/milliliter, wherein the growth medium comprises glucose at a concentration from about 0.1 to about 20 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at a concentration from about 0.1 to about 20 grams/liter; and ii) allowing the mammalian cells to propagate resulting in an inoculum comprising from about 1 to about 50 million cells/milliliter; and b) a production phase comprising: i) inoculating a fed-batch bioreactor, containing a cell culture production medium, with the inoculum comprising from about 3 million to about 20 million cells/milliliter, wherein the production medium comprises glucose at an initial concentration from out 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter, such that adalimumab is produced; ii) maintaining the glucose in the production medium at a concentration from about 0.1 to about 0.9 grams/liter; iii) optionally, maintaining the at least one other hexose in the production medium at a concentration from about 0.1 to about 20 grams/liter; iv) optionally, supplementing the production medium with at least one additional feed absent glucose; and v) harvesting the mammalian cell culture at a cell viability from about 20% to about 100%. 8. The method of claim 7 wherein the cell viability is from about 50% to about 80%. 9. A method of producing adalimumab in a mammalian cell culture comprising: a) inoculating a fed-batch bioreactor containing a cell culture production medium with mammalian cells comprising a nucleic acid encoding adalimumab or a fragment thereof wherein the mammalian cells are at a concentration from about 0.1 million to about 2 million cells/milliliter, wherein the production medium comprises glucose at an initial concentration from about 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter, such that adalimumab is produced; b) maintaining the glucose in the production medium at a concentration from about 0.1 to about 0.9 grams/liter; c) optionally, maintaining the at least one other hexose in the production medium at a concentration from about 0.1 to about 20 grams/liter; d) optionally, supplementing the production medium with at least one additional feed absent glucose; and e) harvesting the mammalian cell culture at a cell viability from about 20% to about 100%. 10. The method of claim 9 wherein the cell viability is from about 50% to about 80%. 11. A method of producing adalimumab in a mammalian cell culture comprising: a) a first cycle comprising: i) inoculating a fed-batch bioreactor containing a first cell culture production medium, with mammalian cells comprising a nucleic acid encoding adalimumab or a fragment thereof wherein the mammalian cells are at a concentration from about 0.1 million to about 2 million cells/milliliter, wherein the first production medium comprises glucose at an initial concentration from about 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter; ii) maintaining the glucose in the first production medium at a concentration from about 0.1 to about 0.9 grams/liter; iii) optionally, maintaining the at least one other hexose in the first production medium at a concentration from about 0.1 to about 20 grams/liter; iv) optionally, supplementing the first production medium with at least one additional feed absent glucose; and v) harvesting 90% mammalian cell culture comprising a cell viability from about 80% to about 100%, and b) a second cycle comprising: i) removing the first production medium from the remaining 10% of the mammalian cell culture; ii) adding a second production medium comprising glucose at an initial concentration from about 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter; iii) maintaining the glucose in the second production medium at a concentration from about 0.1 to about 0.9 grams/liter; iv) optionally, maintaining the at least one other hexose in the second production medium at a concentration from about 0.1 to about 20 grams/liter; v) optionally, supplementing the second production medium with at least one additional feed absent glucose; and vi) harvesting 90% mammalian cell culture comprising a cell viability from about 80% to about 100%; and c) optionally, repeating the first and second cycle, such that adalimumab is produced. 12. A method of producing adalimumab in a mammalian cell culture comprising: a) a growth phase comprising: i) inoculating a perfusion bioreactor containing a cell culture growth medium with mammalian cells comprising a nucleic acid encoding adalimumab or a fragment thereof wherein the mammalian cells are at a concentration from about 0.1 million to about 5 million cells/milliliter, wherein the growth medium comprises glucose at an initial concentration from about 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter; and; ii) allowing the mammalian cells to propagate resulting in the mammalian cell culture comprising from about 10 to about 40 million cells/milliliter; and b) a production phase comprising: i) removing the growth medium; ii) adding a production medium comprising glucose at a concentration from about 0.1 to about 0.9 grams/liter and at least one other hexose selected from the group consisting of galactose, mannose, fructose, maltose, or a combination thereof at an initial concentration from about 0.1 to about 20 grams/liter; iii) maintaining the glucose in the production medium at a concentration from about 0.1 to about 0.9 grams/liter; iv) optionally, maintaining the at least one other hexose in the production medium at a concentration from about 0.1 to about 20 grams/liter; v) optionally, supplementing the production medium with at least one additional feed absent glucose; and vi) harvesting the mammalian cell culture comprising a cell viability from about 20% to about 100%, such that adalimumab is produced. 13. The method of claim 12 wherein the cell viability is from about 50% to about 80%. 14. The method of claim 7, 9, 11, or 12 wherein the mammalian cells are Chinese Hamster Ovary Cells.

Details for Patent 10,435,464

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2034-09-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2034-09-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2034-09-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	09/23/2014		2034-09-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	11/23/2015		2034-09-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	03/09/2016		2034-09-05