Claims for Patent: 9,234,032

Summary for Patent: 9,234,032

Title:	Fed-batch methods for producing adalimumab
Abstract:	The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.
Inventor(s):	Pla; Itzcoatl A. (Worcester, MA), Matuck; Joseph G. (Worcester, MA), Fann; John C. (Shrewsbury, MA), Schulz; Christof (Ayer, MA), Roy; Nichole A. (Worcester, MA), Bruton; David F. (Enfield, CT), McIntire; James (Castro Valley, CA), Chang; Yu-hsiang D. (Solana Beach, CA), Seewoester; Thomas (Simi Valley, CA)
Assignee:	AbbVie Inc. (North Chicago, IL)
Application Number:	14/157,460
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 9,234,032
Patent Claims:	1. A fed-batch method for producing adalimumab in mammalian cells in culture, wherein said mammalian cells express said adalimumab, the method comprising: culturing the mammalian cells in a cell culture production medium in large scale at a first temperature; and then reducing the temperature under which the mammalian cells are cultured to a second lower temperature during adalimumab production, wherein said adalimumab is produced at a titer of at least 1.3 g/L in said cell culture and said adalimumab is further purified by a process including Protein A affinity chromatography. 2. The fed-batch method according to claim 1, wherein the first temperature is 35.degree. C., 37.degree. C., or 38.degree. C. 3. The fed-batch method according to claim 1, wherein the second temperature is 32.degree. C. or 33.degree. C. 4. The fed-batch method according to claim 1, wherein the first temperature is 37.degree. C. and the second temperature is 33.degree. C. 5. The fed-batch method according to claim 1, wherein the first temperature is 37.degree. C. and the second temperature is 32.degree. C. 6. The fed-batch method according to claim 1, wherein the first temperature is 35.degree. C. and the second temperature is 32.degree. C. 7. The fed-batch method according to claim 1, wherein the cell culture production medium comprises at least two different non-animal-based hydrolysates. 8. The fed-batch method according to claim 7, wherein the non-animal-based hydrolysates comprise a plant-based hydrolysate and a yeast-based hydrolysate. 9. The fed-batch method according to claim 8, wherein the plant-based hydrolysate is a soy-based hydrolysate. 10. The fed-batch method according to claim 1, wherein the mammalian cells are Chinese Hamster Ovary (CHO) cells and the cell culture production medium is supplemented with a 25.times. PFCHO solution and a 33.times. solution of a combination of the plant-based hydrolysate Phytone and yeast-based hydrolysate Yeastolate. 11. The fed-batch method according to claim 1, wherein the cell culture production medium maintains a glucose concentration of at least 2 g/L. 12. The fed-batch method according to claim 1, wherein the cell culture production medium comprises: a) a modified basal medium; b) 8 to 10 ml/kg or 110 to 130 mg/L ferric citrate; c) 4 to 8 mL/kg or 10 to 14 mg/kg recombinant human insulin; d) 5 to 9 g/kg anhydrous glucose; e) 0.1 to 1 g/kg L-glutamine; f) 1 to 3 g/kg sodium bicarbonate; g) 1 to 3 g/kg HEPES; h) 2 to 3 g/kg NaCl; i) 0.1 to 2 g/kg Pluronic F-68; j) 0.01 to 0.1 g/kg NaH.sub.2PO.sub.4--H.sub.2O; k) 0.1 to 0.1 g/kg Na.sub.2HPO.sub.4-7H.sub.2O; l) 8 to 12 g/kg yeast-based hydrolysate; and m) 6 to 8 g/kg plant-based hydrolysate. 13. The fed-batch method according to claim 1, wherein said adalimumab is produced at a titer of at least 2 g/L in said cell culture. 14. The fed-batch method according to claim 13, wherein said adalimumab is produced at a titer of at least 4 g/L in said cell culture. 15. The fed-batch method according to claim 1, wherein the temperature is reduced after at least 3 days of culturing the mammalian cells at the first temperature. 16. The fed-batch method according to claim 1, further comprising culturing the mammalian cells in the cell culture production medium in large scale at a starting pH; and then reducing the pH during adalimumab production phase to a value that is less than the starting pH. 17. The fed-batch method according to claim 16, wherein the starting pH is 8 or less and the reduced pH is 6.5 to 7.0. 18. The fed-batch method according to claim 17, wherein the starting pH is 7.1 and the reduced pH is 6.9. 19. The fed-batch method according to claim 18, wherein the pH is adjusted within the first 72 hours of culturing. 20. The fed-batch method according to claim 17, wherein the first temperature is 35.degree. C., 37.degree. C., or 38.degree. C., and the second temperature is 32.degree. C. or 33.degree. C. 21. The fed-batch method according to claim 20, wherein the mammalian cells expressing adalimumab are cultured at a viable cell density of between 2.0.times.10.sup.6 cells/mL and 5.0.times.10.sup.6 cells/mL. 22. The fed-batch method according to claim 20, wherein the mammalian cells expressing adalimumab are cultured at a viable cell density of between 3.5.times.10.sup.6 cells/mL and 5.0.times.10.sup.6 cells/mL. 23. The fed-batch method according to claim 17, further comprising (1) monitoring the glucose concentration in the cell culture production medium and (2) maintaining the glucose concentration in the cell culture production medium at a concentration of at least 2 g/L, or adding glucose to the cell culture production medium when the glucose concentration in the cell culture production medium decreases to below 2 g/L. 24. The fed-batch method according to claim 17, further comprising (1) monitoring the glucose concentration in the cell culture production medium and (2) maintaining the glucose concentration in the cell culture production medium at a concentration of at least 2 g/L but no greater than 7 g/L. 25. The fed-batch method according to claim 17, further comprising (1) monitoring the glucose concentration in the cell culture production medium and (2) maintaining the glucose concentration in the cell culture production medium at a concentration of at least 2 g/L but no greater than 5 g/L. 26. The method according to claim 24, wherein the osmolarity of the cell culture production medium is 440 mOsm or less, and wherein the first temperature is 35.degree. C., 37.degree. C., or 38.degree. C., and the second temperature is 32.degree. C. or 33.degree. C. 27. The method according to claim 26, wherein the cell culture production medium is maintained at between 20-65% dissolved oxygen. 28. The method according to claim 26, wherein the cell culture production medium is maintained at about 30% dissolved oxygen. 29. The method according to claim 26, wherein the large-scale cell culture is greater than about 10 L. 30. The method according to claim 26, wherein the large-scale cell culture is 13 L. 31. The method according to claim 26, wherein the mammalian cells are Chinese Hamster Ovary (CHO) cells.

Details for Patent 9,234,032

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2026-09-13
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2026-09-13
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2026-09-13
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	09/23/2014		2026-09-13
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	11/23/2015		2026-09-13