Claims for Patent: 9,045,536

Summary for Patent: 9,045,536

Title:	Cell line 3M
Abstract:	The present invention provides, inter alia, an isolated cell line, 3M as well as methods for making such a cell line and methods of using such a cell line, e.g., to produce a protein such as an immunoglobulin.
Inventor(s):	Merchant; Ankit A. (Old Bridge, NJ), Tsao; Yung-Shyeng (Summit, NJ)
Assignee:	MERCK SHARP & DOHME CORP. (Rahway, NJ)
Application Number:	13/517,901
Patent Claims:	1. An isolated Chinese hamster ovary cell deposited at the American Type Culture Collection under deposit number PTA-10481. 2. The cell of claim 1 in an aqueous liquid cell culture medium. 3. The cell of claim 1 in a vessel. 4. The cell of claim 1 which comprises a vector. 5. The cell of claim 4 wherein the vector comprises a polynucleotide encoding one or more proteins. 6. The cell of claim 1 comprising a protein that is an immunoglobulin comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-8, 21, 22, 42, 43, 58-69, 79, 80, 81, 85, 89, 93 and 101 or a mature fragment thereof or an immunoglobulin comprising one or more CDRs from said immunoglobulin; optionally linked to an immunoglobulin constant chain; or one or both chains of etanercept. 7. The cell of claim 1 comprising a protein that is an immunoglobulin chain comprising: TABLE-US-00030 (1) CDR-L1: (SEQ ID NO: 9) KASKKVTIFGSISALH, CDR-L2: (SEQ ID NO: 10) NGAKLES, and CDR-L3: (SEQ ID NO: 11) LQNKEVPYT; (2) CDR-H1: (SEQ ID NO: 12) SYGIT, CDR-H2: (SEQ ID NO: 13) ENYPRSGNTYYNEKFKG, and CDR-H3: (SEQ ID NO: 14) CEFISTVVAPYYYALDY or (SEQ ID NO: 15) SEFISTVVAPYYYALDY or (SEQ ID NO: 16) AEFISTVVAPYYYALDY or (SEQ ID NO: 17) VEFISTVVAPYYYALDY or (SEQ ID NO: 18) SEFISTVMAPYYYALDY or (SEQ ID NO: 19) SEFTSTVVAPYYYALDY; (3) CDRH1: (SEQ ID NO: 23) Gly Phe Thr Phe Ser Ser Tyr Thr Met Ser, CDRH2: (SEQ ID NO: 24) Thr Ile Ser Ser Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys Gly, and CDRH3: (SEQ ID NO: 25) Asp Asn His Ala Tyr Asp Arg Gly Pro Phe Phe Asp Tyr; (4) CDRL1: (SEQ ID NO: 26) Lys Ser Ser Gln Asn Leu Phe Tyr Arg Ser Asn Gln Lys Asn His Leu Ala, CDRL2: (SEQ ID NO: 27) Trp Thr Ser Thr Arg Glu Ser, and CDRL3: (SEQ ID NO: 28) Gln Gln Tyr Tyr Ser Tyr Pro Pro Thr; (5) CDRH1: (SEQ ID NO: 29) Ala Tyr Gly Met Asp, CDRH2: (SEQ ID NO: 30) Ser Ile Ser Pro Ser Gly Gly Arg Thr Lys Tyr Ala Asp Ser Val Lys Gly, and CDRH3: (SEQ ID NO: 31) Asp Leu Gly Gly Gly Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val; (6) CDRL1: (SEQ ID NO: 32) Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser, CDRL2: (SEQ ID NO: 33) Glu Val Ser Asn Arg Pro Ser, and CDRL3: (SEQ ID NO: 34) Ser Ser Tyr Thr Ser Ser Ser Thr Leu Phe Tyr Val; (7) CDRH1: (SEQ ID NO: 35) GKTFWSWGIN, CDRH2: (SEQ ID NO: 36) YIYIGTGYTEPNPKYKG, and CDRH3: (SEQ ID NO: 37) IGGYYGNFAD or (SEQ ID NO: 38) IGGYYGNFDQ; (8) CDRL1: (SEQ ID NO: 39) RSSQSLLISGGNTYLN, CDRL2: (SEQ ID NO: 40) LVSKLDQ, and CDRL3: (SEQ ID NO: 41) WQGTYFPLT; (9) CDRH1: (SEQ ID NO: 44) TYWMH or (SEQ ID NO: 45) TYWMH, CDRH2: (SEQ ID NO: 46) EINPTNGHTNYNEKFKS, (SEQ ID NO: 47) EINPTNGHTNYNPSFQG, or (SEQ ID NO: 48) EINPTNGHTNYNQKFQG, and CDRH3: (SEQ ID NO: 49) NYVGSIFDY or (SEQ ID NO: 50) NYVGSIFDY; (10) CDRL1: (SEQ ID NO: 51) KASENVVSYVS or (SEQ ID NO: 52) KASENVVSYVS, CDRL2: (SEQ ID NO: 53) GASNRNT, (SEQ ID NO: 54) GASNRNT or (SEQ ID NO: 55) GASNRES, and CDRL3: (SEQ ID NO: 56) GQSYNYPYT or (SEQ ID NO: 57) GQSYNYPYT; (11) CDRH1: (SEQ ID NO: 82) GYTFSSYGMY; CDRH2: (SEQ ID NO: 83) WIDPGSGGTKYNEKFKG; and CDRH3: (SEQ ID NO: 84) ERYGYYFDY; (12) CDRL1: (SEQ ID NO: 86) RASQYVGSYLN; CDRL2: (SEQ ID NO: 87) DASNRAT; and CDRL3: (SEQ ID NO: 88) QVWDSSPPVV; (13) CDRL1: (SEQ ID NO: 90) RASQDVSRYLT; CDRL2: (SEQ ID NO: 91) AASSLQS; and CDRL3: (SEQ ID NO: 92) QAYDYSLSGYV; (14) CDRH1: (SEQ ID NO: 94) GYTFSSYWMH; CDRH2: (SEQ ID NO: 95) RIDPYNGGTKYNEKFKG; and CDRH3: (SEQ ID NO: 96) YGYYLGSYAMDY; (15) CDRH1: (SEQ ID NO: 98) GYTFTDYYMN; CDHR2: (SEQ ID NO: 99) DINPNNGGAIYNQKFKG; and CDRH3: (SEQ ID NO: 100) GIITEIAEDF; or (16) CDRL1: (SEQ ID NO: 102) KASQNVGTNVV; CDRL2: (SEQ ID NO: 103) SASYRYS; and CDRL3: (SEQ ID NO: 104) QQYKTYPYT; optionally linked to an immunoglobulin constant chain. 8. A method for making one or more polypeptides; or antibodies or antigen-binding fragments thereof that comprise said polypeptides; comprising introducing one or more polynucleotides encoding said polypeptides into the cell of claim 1 and culturing the cell in conditions under which the polypeptides are produced. 9. The method of claim 8 further comprising isolating the polypeptide. 10. The method of claim 8 wherein the polypeptide is an immunoglobulin. 11. The method of claim 10 wherein the immunoglobulin comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-8, 21, 22, 42, 43, 58-69, 79, 80, 81, 85, 89, 93 and 101 or a mature fragment thereof or an immunoglobulin comprising one or more CDRs from said immunoglobulin; optionally linked to an immunoglobulin constant chain. 12. The method of claim 10 wherein the immunoglobulin comprises: TABLE-US-00031 (1) CDR-L1: (SEQ ID NO: 9) KASKKVTIFGSISALH, CDR-L2: (SEQ ID NO: 10) NGAKLES, and CDR-L3: (SEQ ID NO: 11) LQNKEVPYT; (2) CDR-H1: (SEQ ID NO: 12) SYGIT, CDR-H2: (SEQ ID NO: 13) ENYPRSGNTYYNEKFKG, and CDR-H3: (SEQ ID NO: 14) CEFISTVVAPYYYALDY or (SEQ ID NO: 15) SEFISTVVAPYYYALDY or (SEQ ID NO: 16) AEFISTVVAPYYYALDY or (SEQ ID NO: 17) VEFISTVVAPYYYALDY or (SEQ ID NO: 18) SEFISTVMAPYYYALDY or (SEQ ID NO: 19) SEFTSTVVAPYYYALDY; (3) CDRH1: (SEQ ID NO: 23) Gly Phe Thr Phe Ser Ser Tyr Thr Met Ser, CDRH2: (SEQ ID NO: 24) Thr Ile Ser Ser Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys Gly, and CDRH3: (SEQ ID NO: 25) Asp Asn His Ala Tyr Asp Arg Gly Pro Phe Phe Asp Tyr; (4) CDRL1: (SEQ ID NO: 26) Lys Ser Ser Gln Asn Leu Phe Tyr Arg Ser Asn Gln Lys Asn His Leu Ala, CDRL2: (SEQ ID NO: 27) Trp Thr Ser Thr Arg Glu Ser, and CDRL3: (SEQ ID NO: 28) Gln Gln Tyr Tyr Ser Tyr Pro Pro Thr; (5) CDRH1: (SEQ ID NO: 29) Ala Tyr Gly Met Asp, CDRH2: (SEQ ID NO: 30) Ser Ile Ser Pro Ser Gly Gly Arg Thr Lys Tyr Ala Asp Ser Val Lys Gly, and CDRH3: (SEQ ID NO: 31) Asp Leu Gly Gly Gly Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val; (6) CDRL1: (SEQ ID NO: 32) Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser, CDRL2: (SEQ ID NO: 33) Glu Val Ser Asn Arg Pro Ser, and CDRL3: (SEQ ID NO: 34) Ser Ser Tyr Thr Ser Ser Ser Thr Leu Phe Tyr Val; (7) CDRH1: (SEQ ID NO: 35) GKTFWSWGIN, CDRH2: (SEQ ID NO: 36) YIYIGTGYTEPNPKYKG, and CDRH3: (SEQ ID NO: 37) IGGYYGNFAD or (SEQ ID NO: 38) IGGYYGNFDQ; (8) CDRL1: (SEQ ID NO: 39) RSSQSLLISGGNTYLN, CDRL2: (SEQ ID NO: 40) LVSKLDQ, and CDRL3: (SEQ ID NO: 41) WQGTYFPLT; (9) CDRH1: (SEQ ID NO: 44) TYWMH or (SEQ ID NO: 45) TYWMH, CDRH2: (SEQ ID NO: 46) EINPTNGHTNYNEKFKS, (SEQ ID NO: 47) EINPTNGHTNYNPSFQG, or (SEQ ID NO: 48) EINPTNGHTNYNQKFQG, and CDRH3: (SEQ ID NO: 49) NYVGSIFDY or (SEQ ID NO: 50) NYVGSIFDY; (10) CDRL1: (SEQ ID NO: 51) KASENVVSYVS or (SEQ ID NO: 52) KASENVVSYVS, CDRL2: (SEQ ID NO: 53) GASNRNT, (SEQ ID NO: 54) GASNRNT or (SEQ ID NO: 55) GASNRES, and CDRL3: (SEQ ID NO: 56) GQSYNYPYT or (SEQ ID NO: 57) GQSYNYPYT; (11) CDRH1: (SEQ ID NO: 82) GYTFSSYGMY; CDRH2: (SEQ ID NO: 83) WIDPGSGGTKYNEKFKG; and CDRH3: (SEQ ID NO: 84) ERYGYYFDY; (12) CDRL1: (SEQ ID NO: 86) RASQYVGSYLN; CDRL2: (SEQ ID NO: 87) DASNRAT; and CDRL3: (SEQ ID NO: 88) QVWDSSPPVV; (13) CDRL1: (SEQ ID NO: 90) RASQDVSRYLT; CDRL2: (SEQ ID NO: 91) AASSLQS; and CDRL3: (SEQ ID NO: 92) QAYDYSLSGYV; (14) CDRH1: (SEQ ID NO: 94) GYTFSSYWMH; CDRH2: (SEQ ID NO: 95) RIDPYNGGTKYNEKFKG; and CDRH3: (SEQ ID NO: 96) YGYYLGSYAMDY; (15) CDRH1: (SEQ ID NO: 98) GYTFTDYYMN; CDHR2: (SEQ ID NO: 99) DINPNNGGAIYNQKFKG; and CDRH3: (SEQ ID NO: 100) GIITEIAEDF; or (16) CDRL1: (SEQ ID NO: 102) KASQNVGTNVV; CDRL2: (SEQ ID NO: 103) SASYRYS; and CDRL3: (SEQ ID NO: 104) QQYKTYPYT; optionally linked to an immunoglobulin constant chain. 13. The method of claim 10 wherein the immunoglobulin is a light chain immunoglobulin linked to a kappa or lambda constant immunoglobulin chain and/or wherein the immunoglobulin is a heavy chain immunoglobulin linked to a gamma-1, gamma-2, gamma-3 or gamma-4 constant immunoglobulin chain. 14. A method for producing an antibody comprising inoculating an initial mammalian cell growth medium, pre-warmed to about 37.degree. C.; which initial medium comprises HEPES, sodium bicarbonate buffers, inorganic salts, non-essential amino acids, recombinant human insulin, trace elements and surfactants; and which does not comprise L-glutamine, antibiotics, antimycotics or animal-derived components; with one or more cells of claim 1 expressing the antibody light chain immunoglobulin and heavy chain immunoglobulin, to a cell density of about 2.5-5.times.10.sup.5 cells/ml; and, adding the following supplements to the medium before, simultaneously with or immediately after said inoculation: soy hydrolysate to a final concentration of about 10 g/liter; and, optionally, an amino acid feed wherein the concentration of the components added by said amino acid feed are approximately those set forth below: TABLE-US-00032 L-arginine: 126.4 mg/liter L-cystine: 34 mg/liter L-histidine: 42 mg/liter L-isoleucine: 52 mg/liter L-leucine: 52 mg/liter L-lysine: 72 mg/liter L-Methionine: 15.2 mg/liter L-phenylalanine: 33 mg/liter L-threonine: 47.6 mg/liter L-tryptophan: 10.2 mg/liter L-tyrosine: 36 mg/liter L-valine: 46.8 mg/liter L-alanine: 8.9 mg/liter L-asparagine: 30 mg/liter L-aspartic acid: 26.6 mg/liter L-glutamic acid: 29.4 mg/liter glycine: 15 mg/liter L-proline: 23 mg/liter L-serine: 21 mg/liter; and, when viable cell density reaches over about 1.2.times.10.sup.6 cells/ml, adding supplement feeds wherein the concentration of the components added by said supplement feeds are approximately those set forth below: TABLE-US-00033 Sodium selenite: 0.01426 mg/liter Adenine sulfate: 1.632 mg/liter Adenosine: 17.6 mg/liter Cytidine: 17.6 mg/liter Guanosine: 17.6 mg/liter Uridine: 17.6 mg/liter Hypoxanthine: 11.8 mg/liter L-citrulline: 12.6 mg/liter L-ornithine-HCl: 25.6 mg/liter Biotin: 0.28 mg/liter Flavin Adenine Dinucleotide: 0.05 mg/liter Folic Acid: 4.6 mg/liter Lipoic Acid: 0.52 mg/liter Niacin: 31.4 mg/liter Pyridoxine HCl: 3 mg/liter Riboflavin: 1.86 mg/liter Thiamine HCl: 16 mg/liter Vitamin E: 0.376 mg/liter Vitamin B12: 3.4 mg/liter Choline Chloride: 50.2 mg/liter Ethanolamine HCl: 4.4 mg/liter i-Inositol: 73.2 mg/liter Thymidine: 7.8 mg/liter Putrescine 2HCl: 0.4 mg/liter Progesterone: 0.015 mg/liter D-Calcium Pantothenate: 23.8 mg/liter L-asparagine: 812 mg/liter L-proline 216 mg/liter L-isoleucine 370 mg/liter L-cysteine-HCl 224 mg/liter L-leucine 332 mg/liter L-threonine 164 mg/liter L-tyrosine 198 mg/liter L-arginine 186 mg/liter L-aspartic acid 71 mg/liter L-glutamic acid 126 mg/liter Glycine 57 mg/liter L-histidine 125 mg/liter L-methionine 132 mg/liter L-tryptophan 99 mg/liter L-lysine 293 mg/liter L-phenylalanine 174 mg/liter L-valine 262 mg/liter L-serine: 260 mg/liter Sodium phosphate monobasic: 288.2 mg/liter Zinc sulfate: 1.08 mg/liter Cupric sulfate: 0.0032 mg/liter Ammonium vanadate: 0.00078 mg/liter Cobalt chloride: 0.0025 mg/liter Nickel dichloride hexahydrate: 0.0004 mg/liter Sodium molybdate dehydrate: 0.00016 mg/liter; and, maintaining glucose concentration in the medium at about 1.5 g/liter and maintaining L-glutamine concentration in the medium at about 150 mg/liter; and during cell growth maintaining O.sub.2 concentration at about 60%; pH at about 6.8.+-.0.02 and temperature at about 36.5.degree. C..+-.0.5.degree. C.; and, optionally, removing the host cells from the medium when cell viability is below about 60%. 15. The method of claim 14 further comprising recovering the culture medium from the cells by disk-stack centrifuging the medium, depth filtering the medium and filtering the medium through a filter with about a 0.2 micron pore size. 16. The method of claim 14 further comprising purifying the immunoglobulins from the medium by column chromatographic fractionation. 17. A master cell bank or working cell bank comprising the cell of claims 1. 18. The cell of claim 1 in a cell freezing medium. 19. The cell of claim 18 wherein the cell freezing medium comprises dimethylsulfoxide. 20. A method for making a Chinese hamster ovary cell comprising adapting CHO-DXB11 cells into animal-component free medium in suspension for 83 days; then subcloning the cells into said medium twice.

Details for Patent 9,045,536

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Immunex Corporation	ENBREL	etanercept	For Injection	103795	11/02/1998		2029-12-23
Immunex Corporation	ENBREL	etanercept	For Injection	103795	05/27/1999		2029-12-23
Immunex Corporation	ENBREL	etanercept	Injection	103795	09/27/2004		2029-12-23
Immunex Corporation	ENBREL	etanercept	Injection	103795	02/01/2007		2029-12-23
Immunex Corporation	ENBREL MINI	etanercept	Injection	103795	09/14/2017		2029-12-23
Immunex Corporation	ENBREL	etanercept	Injection	103795			2029-12-23
Immunex Corporation	ENBREL	etanercept	Injection	103795	03/05/2020		2029-12-23