Claims for Patent: 9,714,293

Summary for Patent: 9,714,293

Title:	Production of proteins in glutamine-free cell culture media
Abstract:	The present invention relates generally to glutamine-free cell culture media supplemented with asparagine. The invention further concerns the production of recombinant proteins, such as antibodies, in asparagine-supplemented glutamine-free mammalian cell culture.
Inventor(s):	Gawlitzek; Martin (Redwood City, CA), Luo; Shun (Irvine, CA), Bevilacqua; Christina Teresa (San Ramon, CA)
Assignee:	GENENTECH, INC. (South San Francisco, CA)
Application Number:	14/670,079
Patent Litigation and PTAB cases:	See patent lawsuits and PTAB cases for patent 9,714,293
Patent Claims:	1. A process for producing a polypeptide in a host cell expressing said polypeptide, comprising culturing the host cell in a production phase of the culture in a glutamine-free production culture medium containing asparagine and aspartic acid, wherein the asparagine is added at a concentration in the range of 7.5 mM to 15 mM and wherein the aspartic acid is added at a concentration in the range of 1 mM to 10 mM. 2. The process of claim 1 further comprising the step of isolating said polypeptide. 3. The process of claim 2 further comprising determining one or more of cell viability, culture longevity, specific productivity and final recombinant protein titer following isolation. 4. The process of claim 3 wherein at least one of the cell viability, culture longevity, specific productivity and final recombinant protein titer is increased relative to the cell viability, culture longevity, specific productivity and final recombinant protein titer in a glutamine-containing production medium of the same composition. 5. The process of claim 1 wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 6. The process of claim 1 wherein the asparagine is added at a concentration of 10 mM. 7. The process of claim 1 wherein the production medium is serum-free. 8. The process of claim 1 wherein the production culture medium comprises one or more ingredients selected from the group consisting of 1) an energy source; 2) essential amino acids; 3) vitamins; 4) free fatty acids; and 5) trace elements. 9. The process of claim 8 wherein the production culture medium additionally comprises one or more ingredients selected from the group consisting of: 1) hormones and other growth factors; 2) salts and buffers; and 3) nucleosides. 10. The process of claim 1 wherein the production phase is a batch or fed batch culture phase. 11. The process of claim 10, wherein the production culture medium comprises one or more ingredients selected from the group consisting of 1) an energy source; 2) essential amino acids; 3) vitamins; 4) free fatty acids; and 5) trace elements. 12. The process of claim 11, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 13. The process of 11, wherein the asparagine is added at a concentration of 10 mM. 14. The process of claim 11, wherein the aspartic acid is added at a concentration of 10 mM. 15. The process of claim 11, wherein the production medium is serum-free. 16. The process of claim 10, wherein the production culture medium additionally comprises one or more ingredients selected from the group consisting of: 1) hormones and other growth factors; 2) salts and buffers; and 3) nucleosides. 17. The process of claim 16, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 18. The process of claim 16, wherein the asparagine is added at a concentration of 10 mM. 19. The process of claim 16, wherein the aspartic acid is added at a concentration of 10 mM. 20. The process of claim 16, wherein the production medium is serum-free. 21. The process of claim 1 wherein said host cell is an eukaryotic host cell. 22. The process of claim 21 wherein said eukaryotic host cell is a mammalian host cell. 23. The process of claim 22, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 24. The process of claim 22, wherein the asparagine is added at a concentration of 10 mM. 25. The process of claim 22, wherein the aspartic acid is added at a concentration of 10 mM. 26. The process of claim 22, wherein the production medium is serum-free. 27. The process of claim 22, wherein the production culture medium comprises one or more ingredients selected from the group consisting of 1) an energy source; 2) essential amino acids; 3) vitamins; 4) free fatty acids; and 5) trace elements. 28. The process of claim 27, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 29. The process of claim 27, wherein the asparagine is added at a concentration of 10 mM. 30. The process of claim 27, wherein the production culture medium additionally comprises one or more ingredients selected from the group consisting of: 1) hormones and other growth factors; 2) salts and buffers; and 3) nucleosides. 31. The process of claim 22, wherein the production culture medium additionally comprises one or more ingredients selected from the group consisting of: 1) hormones and other growth factors; 2) salts and buffers; and 3) nucleosides. 32. The process of claim 31, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 33. The process of claim 31, wherein the asparagine is added at a concentration of 10 mM. 34. The process of claim 31, wherein the aspartic acid is added at a concentration of 10 mM. 35. The process of claim 31, wherein the production medium is serum-free. 36. The process of claim 22, wherein the production phase is a batch or fed batch culture phase. 37. The process of claim 36, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 38. The process of claim 36, wherein the asparagine is added at a concentration of 10 mM. 39. The process of claim 36, wherein the aspartic acid is added at a concentration of 10 mM. 40. The process of claim 36, wherein the production medium is serum-free. 41. The process of claim 22 wherein said mammalian host cell is a Chinese Hamster Ovary (CHO) cell. 42. The process of claim 41 wherein the mammalian host cell is a dhfr- CHO cell. 43. The process of claim 41, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 44. The process of claim 41, wherein the asparagine is added at a concentration of 10 mM. 45. The process of claim 41, wherein the aspartic acid is added at a concentration of 10 mM. 46. The process of claim 41, wherein the production medium is serum-free. 47. The process of claim 41, wherein the production culture medium comprises one or more ingredients selected from the group consisting of 1) an energy source; 2) essential amino acids; 3) vitamins; 4) free fatty acids; and 5) trace elements. 48. The process of claim 47, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 49. The process of claim 47, wherein the asparagine is added at a concentration of 10 mM. 50. The process of claim 47, wherein the production culture medium additionally comprises one or more ingredients selected from the group consisting of: 1) hormones and other growth factors; 2) salts and buffers; and 3) nucleosides. 51. The process of claim 41, wherein the production culture medium additionally comprises one or more ingredients selected from the group consisting of: 1) hormones and other growth factors; 2) salts and buffers; and 3) nucleosides. 52. The process of claim 51, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 53. The process of claim 51, wherein the asparagine is added at a concentration of 10 mM. 54. The process of claim 51, wherein the aspartic acid is added at a concentration of 10 mM. 55. The process of claim 51, wherein the production medium is serum-free. 56. The process of claim 41, wherein the production phase is a batch or fed batch culture phase. 57. The process of claim 56, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 58. The process of claim 56, wherein the asparagine is added at a concentration of 10 mM. 59. The process of claim 56, wherein the aspartic acid is added at a concentration of 10 mM. 60. The process of claim 56, wherein the production medium is serum-free. 61. The process of claim 56, wherein the production culture medium comprises one or more ingredients selected from the group consisting of 1) an energy source; 2) essential amino acids; 3) vitamins; 4) free fatty acids; and 5) trace elements. 62. The process of claim 61, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 63. The process of claim 61, wherein the asparagine is added at a concentration of 10 mM. 64. The process of claim 61, wherein the aspartic acid is added at a concentration of 10 mM. 65. The process of claim 1 wherein the polypeptide is a mammalian glycoprotein. 66. The process of claim 1 wherein the polypeptide is selected from the group consisting of antibodies, antibody fragments, and immunoadhesins. 67. The process of claim 66 wherein said antibody fragment is selected from the group consisting of Fab, Fab', F(ab')2, scFv, (scFv)2, dAb, complementarity determining region (CDR) fragments, linear antibodies, single-chain antibody molecules, minibodies, diabodies, and multispecific antibodies formed from antibody fragments. 68. The process of claim 66 wherein the antibody or antibody fragment is chimeric, humanized or human. 69. The process of claim 66 wherein said antibody or antibody fragment is a therapeutic antibody or a biologically functional fragment thereof. 70. The process of claim 69 wherein said therapeutic antibody is selected from the group consisting of anti-HER2 antibodies; anti-CD20 antibodies; anti-IL-8 antibodies; anti-VEGF antibodies; anti-CD40 antibodies; anti-CD 11 a antibodies; anti-CD 18 antibodies; anti-IgE antibodies; anti-Apo-2 receptor antibodies; anti-Tissue Factor (TF) antibodies; anti-human .alpha.4.beta.7 integrin antibodies; anti-EGFR antibodies; anti-CD3 antibodies; anti-CD25 antibodies; anti-CD4 antibodies; anti-CD52 antibodies; anti-Fc receptor antibodies; anti-carcinoembryonic antigen (CEA) antibodies; antibodies directed against breast epithelial cells; antibodies that bind to colon carcinoma cells; anti-CD38 antibodies; anti-CD33 antibodies; anti-CD22 antibodies; anti-EpCAM antibodies; anti-GpIIb/IIIa antibodies; anti-RSV antibodies; anti-CMV antibodies; anti-HIV antibodies; anti-hepatitis antibodies; anti-CA 125 antibodies; anti-.alpha.v.beta.3 antibodies; anti-human renal cell carcinoma antibodies; anti-human 17-1A antibodies; anti-human colorectal tumor antibodies; anti-human melanoma antibody R24 directed against GD3 ganglioside; anti-human squamous-cell carcinoma; and anti-human leukocyte antigen (HLA) antibodies, and anti-HLA DR antibodies. 71. The process of claim 69 wherein said therapeutic antibody is an antibody binding to a HER receptor, VEGF, IgE, CD20, CD11a, CD40, BR3 or DR5. 72. The process of claim 71, wherein the therapeutic antibody is selected from the group consisting of bevacizumab, rituximab, and trastuzumab. 73. The process of claim 72, wherein the asparagine is added at a concentration in the range of 7.5 mM to 10 mM. 74. The process of claim 72, wherein the asparagine is added at a concentration of 10 mM. 75. The process of claim 72, wherein the aspartic acid is added at a concentration of 10 mM. 76. The process of claim 72, wherein the production medium is serum-free. 77. The process of claim 1 wherein said polypeptide is a therapeutic polypeptide. 78. The process of claim 77 wherein said therapeutic polypeptide is selected from the group consisting of a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clotting factors such as factor VIIIC, factor IX, tissue factor, and von Willebrands factor; anti-clotting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha and -beta; enkephalinase; RANTES (regulated on activation normally T-cell expressed and secreted); human macrophage inflammatory protein (MIP-1-alpha); a serum albumin such as human serum albumin; Muellerian-inhibiting substance; relaxin A-chain; relaxin B-chain; prorelaxin; mouse gonadotropin-associated peptide; a microbial protein, such as beta-lactamase; DNase; IgE; a cytotoxic T-lymphocyte associated antigen (CTLA), such as CTLA-4; inhibin; activin; vascular endothelial growth factor (VEGF); receptors for hormones or growth factors; Protein A or D; rheumatoid factors; a neurotrophic factor such as bone-derived neurotrophic factor (BDNF), neurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6), or a nerve growth factor such as NGF-.beta.; platelet-derived growth factor (PDGF); fibroblast growth factor such as aFGF and bFGF; epidermal growth factor (EGF); transforming growth factor (TGF) such as TGF-alpha and TGF-beta, including TGF-.beta.1, TGF-.beta.2, TGF-.beta.3, TGF-.beta.4, or TGF-.beta.5; insulin-like growth factor-I and -II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding proteins; CD proteins such as CD3, CD4, CD8, CD19, CD20, CD34, and CD40; erythropoietin; osteoinductive factors; immunotoxins; a bone morphogenetic protein (BMP); an interferon such as interferon-alpha, -beta, and -gamma; colony stimulating factors (CSFs), e.g., M-CSF, GM-CSF, and G-CSF; interleukins (ILs), e.g., IL-1 to IL-10; superoxide dismutase; T-cell receptors; surface membrane proteins; decay accelerating factor; viral antigen such as, for example, a portion of the AIDS envelope; transport proteins; homing receptors; addressins; regulatory proteins; integrins such as CD11a, CD11b, CD11c, CD18, an ICAM, VLA-4 and VCAM; a tumor associated antigen such as HER2, HER3 or HER4 receptor; and fragments of said polypeptides.

Details for Patent 9,714,293

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Microbix Biosystems Inc.	KINLYTIC	urokinase	For Injection	021846	01/16/1978		2029-08-11
Grifols Therapeutics Llc	ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5	albumin (human)	For Injection	101138	10/21/1942		2029-08-11
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		2029-08-11
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		2029-08-11
Grifols Biologicals Llc	ALBUTEIN	albumin (human)	Injection	102478	08/15/1978		2029-08-11