CN101418330A - 适于nso细胞大规模培养及抗体生产的无蛋白培养基 - Google Patents
适于nso细胞大规模培养及抗体生产的无蛋白培养基 Download PDFInfo
- Publication number
- CN101418330A CN101418330A CNA2008100393058A CN200810039305A CN101418330A CN 101418330 A CN101418330 A CN 101418330A CN A2008100393058 A CNA2008100393058 A CN A2008100393058A CN 200810039305 A CN200810039305 A CN 200810039305A CN 101418330 A CN101418330 A CN 101418330A
- Authority
- CN
- China
- Prior art keywords
- vitamin
- protein
- free medium
- acid
- cultivation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108090000623 proteins and genes Proteins 0.000 title abstract description 9
- 102000004169 proteins and genes Human genes 0.000 title abstract description 8
- 238000004519 manufacturing process Methods 0.000 title abstract description 5
- 239000001963 growth medium Substances 0.000 title description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 30
- 229940088594 vitamin Drugs 0.000 claims abstract description 26
- 229930003231 vitamin Natural products 0.000 claims abstract description 26
- 235000013343 vitamin Nutrition 0.000 claims abstract description 26
- 239000011782 vitamin Substances 0.000 claims abstract description 26
- 150000003722 vitamin derivatives Chemical class 0.000 claims abstract description 26
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 claims abstract description 18
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 claims abstract description 18
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 claims abstract description 16
- 239000000203 mixture Substances 0.000 claims abstract description 12
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 claims abstract description 9
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 claims abstract description 9
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229960003531 phenolsulfonphthalein Drugs 0.000 claims abstract description 9
- 229960003080 taurine Drugs 0.000 claims abstract description 9
- 229940104230 thymidine Drugs 0.000 claims abstract description 9
- 239000000186 progesterone Substances 0.000 claims abstract description 8
- 229960003387 progesterone Drugs 0.000 claims abstract description 8
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 claims abstract description 7
- 150000001413 amino acids Chemical class 0.000 claims abstract description 7
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229920001993 poloxamer 188 Polymers 0.000 claims abstract description 7
- 150000003839 salts Chemical class 0.000 claims abstract description 7
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims abstract description 7
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims abstract description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 5
- 239000008103 glucose Substances 0.000 claims abstract description 5
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 4
- 229960004106 citric acid Drugs 0.000 claims abstract description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 14
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 claims description 14
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical group [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 14
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 14
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 14
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims description 12
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 10
- 229930182817 methionine Natural products 0.000 claims description 10
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 claims description 10
- 239000011734 sodium Substances 0.000 claims description 10
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 9
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 9
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 9
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 9
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 9
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 9
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 8
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 claims description 8
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical group [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 8
- 229960004232 linoleic acid Drugs 0.000 claims description 8
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical group [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 7
- 235000019743 Choline chloride Nutrition 0.000 claims description 7
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 claims description 7
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 7
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 7
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 7
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 claims description 7
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 7
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 7
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims description 7
- -1 VITMAIN B1 Chemical compound 0.000 claims description 7
- AGBQKNBQESQNJD-UHFFFAOYSA-N alpha-Lipoic acid Natural products OC(=O)CCCCC1CCSS1 AGBQKNBQESQNJD-UHFFFAOYSA-N 0.000 claims description 7
- FAPWYRCQGJNNSJ-UBKPKTQASA-L calcium D-pantothenic acid Chemical compound [Ca+2].OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O.OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O FAPWYRCQGJNNSJ-UBKPKTQASA-L 0.000 claims description 7
- 239000001110 calcium chloride Substances 0.000 claims description 7
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 7
- 235000012000 cholesterol Nutrition 0.000 claims description 7
- 229940107161 cholesterol Drugs 0.000 claims description 7
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims description 7
- 229960003178 choline chloride Drugs 0.000 claims description 7
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 7
- 229960000304 folic acid Drugs 0.000 claims description 7
- 235000019152 folic acid Nutrition 0.000 claims description 7
- 239000011724 folic acid Substances 0.000 claims description 7
- 229960002885 histidine Drugs 0.000 claims description 7
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims description 7
- 235000019136 lipoic acid Nutrition 0.000 claims description 7
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 7
- 229960003966 nicotinamide Drugs 0.000 claims description 7
- 235000005152 nicotinamide Nutrition 0.000 claims description 7
- 239000011570 nicotinamide Substances 0.000 claims description 7
- 229960005190 phenylalanine Drugs 0.000 claims description 7
- 229960002429 proline Drugs 0.000 claims description 7
- 235000019192 riboflavin Nutrition 0.000 claims description 7
- 229960002477 riboflavin Drugs 0.000 claims description 7
- 239000002151 riboflavin Substances 0.000 claims description 7
- 229910052708 sodium Inorganic materials 0.000 claims description 7
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 7
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 7
- 235000002639 sodium chloride Nutrition 0.000 claims description 7
- 239000011781 sodium selenite Substances 0.000 claims description 7
- 235000015921 sodium selenite Nutrition 0.000 claims description 7
- 229960001471 sodium selenite Drugs 0.000 claims description 7
- 229960002663 thioctic acid Drugs 0.000 claims description 7
- 229960004799 tryptophan Drugs 0.000 claims description 7
- 229960004441 tyrosine Drugs 0.000 claims description 7
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 7
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 7
- 229960001763 zinc sulfate Drugs 0.000 claims description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 6
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 6
- 229940024606 amino acid Drugs 0.000 claims description 6
- 235000001014 amino acid Nutrition 0.000 claims description 6
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 6
- 239000011790 ferrous sulphate Substances 0.000 claims description 6
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 6
- 229940088597 hormone Drugs 0.000 claims description 6
- 239000005556 hormone Substances 0.000 claims description 6
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 6
- 150000002632 lipids Chemical class 0.000 claims description 6
- 239000011573 trace mineral Substances 0.000 claims description 6
- 235000013619 trace mineral Nutrition 0.000 claims description 6
- AGBQKNBQESQNJD-SSDOTTSWSA-N (R)-lipoic acid Chemical compound OC(=O)CCCC[C@@H]1CCSS1 AGBQKNBQESQNJD-SSDOTTSWSA-N 0.000 claims description 5
- KGWDUNBJIMUFAP-KVVVOXFISA-N Ethanolamine Oleate Chemical compound NCCO.CCCCCCCC\C=C/CCCCCCCC(O)=O KGWDUNBJIMUFAP-KVVVOXFISA-N 0.000 claims description 5
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 claims description 5
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 5
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 5
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 5
- XUYPXLNMDZIRQH-LURJTMIESA-N N-acetyl-L-methionine Chemical compound CSCC[C@@H](C(O)=O)NC(C)=O XUYPXLNMDZIRQH-LURJTMIESA-N 0.000 claims description 5
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 5
- 239000005700 Putrescine Substances 0.000 claims description 5
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 5
- 239000006035 Tryptophane Substances 0.000 claims description 5
- 229930003756 Vitamin B7 Natural products 0.000 claims description 5
- 229960001230 asparagine Drugs 0.000 claims description 5
- 239000000872 buffer Substances 0.000 claims description 5
- 229960002079 calcium pantothenate Drugs 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 5
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 5
- 235000019800 disodium phosphate Nutrition 0.000 claims description 5
- 229960002989 glutamic acid Drugs 0.000 claims description 5
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 5
- 229960000367 inositol Drugs 0.000 claims description 5
- MVFCKEFYUDZOCX-UHFFFAOYSA-N iron(2+);dinitrate Chemical compound [Fe+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O MVFCKEFYUDZOCX-UHFFFAOYSA-N 0.000 claims description 5
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 5
- 229960000310 isoleucine Drugs 0.000 claims description 5
- 229940045641 monobasic sodium phosphate Drugs 0.000 claims description 5
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 5
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims description 5
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 5
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 5
- 235000011912 vitamin B7 Nutrition 0.000 claims description 5
- 239000011735 vitamin B7 Substances 0.000 claims description 5
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims description 4
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 4
- 239000004475 Arginine Substances 0.000 claims description 4
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims description 4
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims description 4
- 229930003779 Vitamin B12 Natural products 0.000 claims description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 4
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 claims description 4
- 229920000570 polyether Polymers 0.000 claims description 4
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 claims description 4
- 235000019163 vitamin B12 Nutrition 0.000 claims description 4
- 239000011715 vitamin B12 Substances 0.000 claims description 4
- LXAHHHIGZXPRKQ-UHFFFAOYSA-N 5-fluoro-2-methylpyridine Chemical compound CC1=CC=C(F)C=N1 LXAHHHIGZXPRKQ-UHFFFAOYSA-N 0.000 claims description 3
- 239000004471 Glycine Substances 0.000 claims description 3
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 3
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 3
- 239000004473 Threonine Substances 0.000 claims description 3
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 3
- 229960005261 aspartic acid Drugs 0.000 claims description 3
- 235000003704 aspartic acid Nutrition 0.000 claims description 3
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 3
- 229960001031 glucose Drugs 0.000 claims description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims description 3
- 229960003136 leucine Drugs 0.000 claims description 3
- 229960001153 serine Drugs 0.000 claims description 3
- 229960002898 threonine Drugs 0.000 claims description 3
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 2
- 229930003427 Vitamin E Natural products 0.000 claims description 2
- CMNWUCGLNTVCSI-UHFFFAOYSA-J [Na+].[Na+].[Na+].[Na+].[Cl-].[O-]P([O-])([O-])=O Chemical compound [Na+].[Na+].[Na+].[Na+].[Cl-].[O-]P([O-])([O-])=O CMNWUCGLNTVCSI-UHFFFAOYSA-J 0.000 claims description 2
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 2
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 claims description 2
- 235000019158 vitamin B6 Nutrition 0.000 claims description 2
- 239000011726 vitamin B6 Substances 0.000 claims description 2
- 235000019154 vitamin C Nutrition 0.000 claims description 2
- 239000011718 vitamin C Substances 0.000 claims description 2
- 235000019165 vitamin E Nutrition 0.000 claims description 2
- 229940046009 vitamin E Drugs 0.000 claims description 2
- 239000011709 vitamin E Substances 0.000 claims description 2
- 229940011671 vitamin b6 Drugs 0.000 claims description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims 2
- 102000004452 Arginase Human genes 0.000 claims 1
- 108700024123 Arginases Proteins 0.000 claims 1
- 230000012010 growth Effects 0.000 abstract description 9
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 5
- 238000000746 purification Methods 0.000 abstract description 4
- 238000000926 separation method Methods 0.000 abstract description 4
- 229930024421 Adenine Natural products 0.000 abstract description 3
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 abstract description 3
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 abstract description 3
- 230000006978 adaptation Effects 0.000 abstract description 3
- 229960000643 adenine Drugs 0.000 abstract description 3
- 230000016784 immunoglobulin production Effects 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 239000000047 product Substances 0.000 abstract 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 abstract 1
- 230000002349 favourable effect Effects 0.000 abstract 1
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 230000007774 longterm Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 29
- 239000002609 medium Substances 0.000 description 26
- 239000012679 serum free medium Substances 0.000 description 13
- 238000011160 research Methods 0.000 description 9
- 239000004615 ingredient Substances 0.000 description 7
- 210000004102 animal cell Anatomy 0.000 description 6
- 238000004113 cell culture Methods 0.000 description 6
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 5
- 229960004452 methionine Drugs 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 102000004877 Insulin Human genes 0.000 description 4
- 108090001061 Insulin Proteins 0.000 description 4
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 4
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 102000002070 Transferrins Human genes 0.000 description 4
- 108010015865 Transferrins Proteins 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 150000002505 iron Chemical class 0.000 description 4
- VCJMYUPGQJHHFU-UHFFFAOYSA-N iron(3+);trinitrate Chemical compound [Fe+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O VCJMYUPGQJHHFU-UHFFFAOYSA-N 0.000 description 4
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 235000009697 arginine Nutrition 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- XDIYNQZUNSSENW-UUBOPVPUSA-N (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O XDIYNQZUNSSENW-UUBOPVPUSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 2
- ZAKOWWREFLAJOT-CEFNRUSXSA-N D-alpha-tocopherylacetate Chemical compound CC(=O)OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-CEFNRUSXSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 2
- 235000019766 L-Lysine Nutrition 0.000 description 2
- FFEARJCKVFRZRR-UHFFFAOYSA-N L-Methionine Natural products CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 2
- 229930064664 L-arginine Natural products 0.000 description 2
- 235000014852 L-arginine Nutrition 0.000 description 2
- 235000013878 L-cysteine Nutrition 0.000 description 2
- 239000004201 L-cysteine Substances 0.000 description 2
- 235000019393 L-cystine Nutrition 0.000 description 2
- 239000004158 L-cystine Substances 0.000 description 2
- 229930195722 L-methionine Natural products 0.000 description 2
- 229930182821 L-proline Natural products 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 229960003767 alanine Drugs 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 2
- GICLSALZHXCILJ-UHFFFAOYSA-N ctk5a5089 Chemical compound NCC(O)=O.NCC(O)=O GICLSALZHXCILJ-UHFFFAOYSA-N 0.000 description 2
- 229960003067 cystine Drugs 0.000 description 2
- ZAKOWWREFLAJOT-UHFFFAOYSA-N d-alpha-Tocopheryl acetate Natural products CC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-UHFFFAOYSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 2
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 2
- 235000020778 linoleic acid Nutrition 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 229940054269 sodium pyruvate Drugs 0.000 description 2
- 229940042585 tocopherol acetate Drugs 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 229960004295 valine Drugs 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 102100022641 Coagulation factor IX Human genes 0.000 description 1
- 108010076282 Factor IX Proteins 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- YEESUBCSWGVPCE-UHFFFAOYSA-N azanylidyneoxidanium iron(2+) pentacyanide Chemical compound [Fe++].[C-]#N.[C-]#N.[C-]#N.[C-]#N.[C-]#N.N#[O+] YEESUBCSWGVPCE-UHFFFAOYSA-N 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 150000001886 cortisols Chemical class 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229960004222 factor ix Drugs 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 229960002743 glutamine Drugs 0.000 description 1
- 229960002449 glycine Drugs 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229960002460 nitroprusside Drugs 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000006180 nutrition needs Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明适于NSO细胞大规模培养及抗体生产的无蛋白培养基为一种化学成分明确的无蛋白培养基,其组合物中包括21种氨基酸、13种维生素、8种盐类、4种脂类、6种微量元素,以及孕酮、碳酸氢钠、Hepes、葡萄糖、柠檬酸、牛磺酸、胸苷、腺嘌呤、次黄嘌呤、酚红、巯基乙醇、Pluronic F-68;本发明的积极效果是:无需适应即可支持NSO细胞的长期传代培养,能良好地支持NSO细胞的正常生长及抗体表达;不含蛋白、各种水解物及动物来源成分,化学成分明确,有利于产物的分离纯化,提高医药生物制品的品质;配制方便,适合大规模生产应用。
Description
【技术领域】
本发明涉及现代生物技术之培养基研发技术领域,具体地说,是一种适于NSO细胞大规模培养及抗体生产的无蛋白培养基。
【技术背景】
现代生物技术是21世纪重点发展的前沿技术之一。其中,利用动物细胞培养技术生产具有重要医用价值的生长因子、疫苗和单抗等已成为医药生物高技术产业的重要组成部分。为完善动物细胞培养技术,人们非常关注对培养动物细胞的培养基的研究,因为,培养基是动物细胞培养技术的基础和关键要素之一。在过去几十年里,随着科学技术的发展,动物细胞培养经历了采用有血清培养基、无血清培养基和无蛋白培养基的几个阶段。
动物细胞培养是生产基因工程产品中的重要环节,研究发现,采用含有血清的细胞培养基会给生产及科研带来许多不利因素:第一,影响细胞生长及产品质量,存在批间差异;第二,易被支原体和病毒污染;第三,成分复杂且不确定,给产物的分离纯化带来极大的困难;第四,血清的价格非常昂贵,其费用通常占到培养基整体费用的50%以上,致使生产成本大幅度提高。因此,血清在动物细胞大规模培养中的使用已受到了限制。
二十世纪50年代初,人们开始了对无血清培养基的研究;80年代后,无血清培养基的研究取得了迅速的发展,多种能支持不同种类细胞生长的无血清培养基见诸报端;80年代末90年代初,具有商业应用价值的无血清培养基纷纷上市,其中有:
Darfler等人开发的CITTL无血清培养基〔以DMEM/F12(1:1)为基础培养基,添加了5mg/L的过氧化氢酶、1.5mg/L的胰岛素、1.5~3.0mg/L的转铁蛋白、2nmol/L睾酮、0.5mg/L的二亚油酰磷脂胆碱和1.5mg/L的β-甘油磷酸〕,该无血清培养基可支持s49细胞的生长,也可用于杂交瘤细胞的培养。Chang等人(1980)用所报道的无血清培养基成功地培养了几株杂交瘤细胞。
Murakami等人开发的无血清培养基〔以DMEM/F12(1:1)为基础,添加了5mg/L胰岛素、2~35mg/L转铁蛋白、20μmol/L乙醇胺和1nmol/L亚硒酸钠等物质〕,这就是现在被广泛使用的DMEM/F12-ITES配方。
Kover和Frank开发的无血清培养基〔以RPMI1640为基础培养基,并添加了10mg/L胰岛素、5mg/L转铁蛋白、20μmol/L乙醇胺、5mg/L亚油酸、1g/L牛血清蛋白、3mg/L抗坏血酸、2μg氢化可的松和12种微量元素〕,这一培养基适合几种细胞的生长。
但是,研究中发现,无血清培养基中添加的蛋白成分价格太贵,且有的成分,如牛血清蛋白,含有很多种杂质,给后期的分离纯化带来较大困难,而且目前的新药审批对此方面的要求也越来越严格。因此,在商用无血清培养基之后,科学家们又开始了对无蛋白培养基的研究,从动物细胞的培养基中去掉蛋白成分。
在对细胞营养需求研究的基础上,美国专利5,045,468公开了一种适合杂交瘤细胞生长的无蛋白培养基,该培养基通过添加亚硝基铁氰化物、EDTA、二氧化硒或亚硒酸钠等化合物有效取代了培养基中的蛋白成分。美国专利5,804,420公开了一种适合BHK细胞表达重组因子VIII的无蛋白培养基,其中添加了嵌段式聚醚F68(Pluronic F-68)、硫酸铜、硫酸亚铁、EDTA、锰、钼、硅、锂、铬等成分。另有报道,Jinyou Zhang等人成功地开发了一种适合NSO细胞生长的无蛋白培养基,但由于其成分复杂、还包含了一种价格昂贵、且化学成分未明确的商业混合脂,因此,不适用NSO细胞的大规模培养及抗体生产。
目前,在无蛋白培养基技术领域,以Gibco、Hyclone、JRH等为代表的商业无血清、无蛋白培养基已相继问世,近年来众多研究所报道的培养基大多采用这些商业无蛋白培养基,它们对某些细胞株具有良好的效果;但是,它们同样也存在一些明显的缺点,将其用于某些研究和大规模细胞培养与抗体生产过程会有不少困难,其主要问题是:第一,价格昂贵,只适合一部分实验室小规模研究,不适合大规模细胞培养及抗体生产;第二,大多成分复杂,并且添加了不确定的化学成分,给科研和生产过程带来困难;第三,有些培养基虽能很好地支持细胞生长,但却使细胞表达产物的能力降低,甚至丧失。
【发明内容】
本发明的目的在于克服现有技术的不足,提供一种适于NSO细胞大规模培养及抗体生产的无蛋白培养基,以适应NSO细胞大规模悬浮培养及抗体生产产业化的需要。
为实现上述目的,本发明采取的技术方案为:
采用氨基酸、维生素、盐类、脂类和小分子激素等物质代替传统无血清培养基中的蛋白质和水解物,采用锌离子、EDTA、铁离子和柠檬酸等代替传统无血清培养基中的胰岛素、转铁蛋白和白蛋白,开发出一种适于NSO细胞大规模培养及抗体生产的无蛋白培养基。
一种适于NSO细胞大规模培养及抗体生产的无蛋白培养基,包括氨基酸、维生素、盐类、脂类、微量元素、激素、缓冲剂及葡萄糖、柠檬酸、牛磺酸、胸苷、腺嘌呤、次黄嘌呤、酚红、巯基乙醇、Pluronic F-68,不含蛋白质及水解物的组合物。
所述的氨基酸为丙氨酸、精氨酸、天冬酰胺、天冬氨酸、半胱氨酸、胱氨酸、谷氨酸、谷胺酰胺、甘氨酸、组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、丝氨酸、苏氨酸、色氨酸、酪氨酸、缬氨酸。
所述的维生素为生物素、维生素B12、氯化胆碱、叶酸、肌醇、烟酰胺、泛酸钙、维生素B6、核黄素、维生素B1、腐胺、维生素C、维生素E。
所述的盐类为氯化钙、氯化镁、硫酸镁、氯化钾、磷酸氢二钠、氯化钠、磷酸二氢钠、丙酮酸钠。
所述的脂类为胆固醇、硫辛酸、亚油酸、乙醇胺。
所述的微量元素为硫酸铜、硫酸亚铁、硫酸锌、硝酸铁、亚硒酸钠、EDTA.2Na。
所述的激素为孕酮。
所述的缓冲剂为碳酸氢钠和Hepes。
所述无蛋白培养基中各种成分的含量为:(mg/L)
丙氨酸(L-Alanine) 80.5~104.45
精氨酸(L-Arginine HCI) 280.5~301.2
天冬酰胺(L-Asparagine.H2O) 150.5~202.5
天冬氨酸(L-Aspartic Acid) 70~106.65
半胱氨酸(L-Cysteine.HCI.H2O) 55.56~67.56
胱氨酸(L-Cystine.2HCI) 57~75.73
谷氨酸(L-Glutamic Acid Sodium) 800~1191
谷胺酰胺(L-Glutamine) 230~292
甘氨酸(Glycine) 57~68.75
组氨酸(L-Histidine.HCI.H2O) 55~73.48
异亮氨酸(L-lsoleucine) 80.34~106.87
亮氨酸(L-Leucine) 83~111.45
赖氨酸(L-Lysine.HCI) 130~163.75
甲硫氨酸(L-Methionine) 25~32.34
苯丙氨酸(L-Phenylalanine 50~68.48
脯氨酸(L-Proline) 66.45~84.50
丝氨酸(L-Serine) 80~100
苏氨酸(L-Threonine) 70~101.05
色氨酸(L-Tryptophan) 15.20~19.22
酪氨酸(L-Tyrosine.2Na.2H2O) 70~108.26
缬氨酸(L-Valine) 80.45~99.65
葡萄糖(D-Glucose) 3500~4320
生物素(Biotin) 0.08~0.17
维生素B12(B12Cobalamin) 1.0~2.0
氯化胆碱(Choline Chloride) 6.9~21
叶酸(FolicAcid) 1.4~4.2
肌醇(I-inositol) 9~36
烟酰胺(Niacinamide) 3.3~13
泛酸钙(D-Calcium Pantothenate) 1.8~7
维生素(B6Pyridoxine Hydrochloride) 0.1~0.5
核黄素(Riboflavin) 0.11~0.46
维生素(B1Thiamine Hydrochloride) 1.1~4.4
胸苷(Thymidine) 0.4~1.6
腺嘌呤(Adenine) 0.2~1
腐胺P(utrescine.2HCl) 0.08~0.4
丙酮酸钠(Sodium Pyruvate) 165~330
氯化钙(Calcium Chloride) 81.6~152
氯化镁(Magnesium chloride) 40.3~74.8
酸镁(Magnesium sulfate) 34.2~63.4
氯化钾(Potassium Chloride) 218~404
磷酸氢二钠(Sodium Phosophate Dibasic)397.39
氯化钠(Sodium Chloride)6999.5
磷酸二氢钠(Sodium Phosphate Monobasic.H2O) 62.5
碳酸氢钠(Sodium Bicarbonate) 2438
Hepes 2000~5000
次黄嘌呤(Na Hypoxanthine) 2.04~8.16
酚红(Phenol Red) 8.1
硫酸铜(Cupric Sulfate.5H2O) 0.004~0.012
硫酸亚铁(Ferrous Sulfate.7H2O) 0.62~0.96
维生素(C Ascorbic acid) 4~10
牛磺酸(Taurine) 20~50
维生素(Eα-Tocopherol acetate) 1~4
胆固醇(Cholesterol) 3.5~6.0
硫辛酸(Lipoic acid) 0.08~0.20
亚油酸(Linoleic acid) 0.06~0.15
硫酸锌(Zinc Sulfate.7H2O) 7.1~14.1
硝酸铁(Ferric Nitrate) 0.85~1.21
EDTA二钠(EDTA.2Na) 1.0~5.2
柠檬酸(Citric acid.H2O) 5.0~20
亚硒酸钠(Na2SeO3) 0.002~0.008
乙醇胺(Ethanolamine) 0~1.53
巯基乙醇(β-mercaptoethnol) 0~6.66
嵌段式聚醚F68(Pluronic F-68) 500~2000
孕酮(Progesterone) 0.003~0.012
本发明所述的无蛋白培养基采用常规的制备方法:将上述组分溶解于无热源超纯水即可配制而成。
本发明所述的无蛋白培养基的使用方法也为常规方法。
本发明适于NSO细胞大规模培养及抗体生产的无蛋白培养基的积极效果是:
(1)能良好地支持NSO细胞的正常生长,其最高细胞密度及抗体产量均超过已知的商业培养基;
(2)完全不含蛋白、蛋白水解物成分,不含动物来源成分,是一种化学成分确定的培养基,有利于产物的分离纯化,提高产品的品质;
(3)支持NSO细胞长期传代培养,无需适应即可使用;
(4)成分明确、配制方便,适合于大规模生产。
【附图说明】
附图1为NSO细胞在本发明无蛋白培养基批培养中生长曲线的坐标图;
附图2为NSO细胞在本发明无蛋白培养基批培养中产物表达曲线的坐标图。
【具体实施方式】
以下通过实施例对本发明进行具体地描述,但本发明不限于以下的实施例。
一种适于NSO细胞大规模培养及抗体生产的无蛋白培养基,包括氨基酸、维生素、盐类、脂类、微量元素、激素、缓冲剂及葡萄糖、柠檬酸、牛磺酸、胸苷、腺嘌呤、次黄嘌呤、酚红、巯基乙醇、Pluronic F-68,不含蛋白质及水解物的组合物,其各种成分的含量为:(mg/L)
丙氨酸(L-Alanine) 104.45
精氨酸(L-Arginine HCI) 301.2
天冬酰胺(L-Asparagine.H2O) 202.5
天冬氨酸(L-Aspartic Acid) 106.65
半胱氨酸(L-Cysteine.HCI.H2O) 67.56
胱氨酸(L-Cystine.2HCI) 75.73
谷氨酸(L-Glutamic Acid Sodium) 1191.00
谷胺酰胺(L-Glutamine) 292.00
甘氨酸(Glycine) 68.75
组氨酸(L-Histidine.HCI.H2O) 73.48
异亮氨酸(L-lsoleucine) 106.87
亮氨酸(L-Leucine) 111.45
赖氨酸(L-Lysine.HCI) 163.75
甲硫氨酸(L-Methionine) 32.34
苯丙氨酸(L-Phenylalanine 68.48
脯氨酸(L-Proline) 84.50
丝氨酸(L-Serine) 100.00
苏氨酸(L-Threonine) 101.05
色氨酸(L-Tryptophan) 19.22
酪氨酸(L-Tyrosine.2Na.2H2O) 108.26
缬氨酸(L-Valine) 99.65
葡萄糖(D-Glucose) 4320.00
生物素(Biotin) 0.11
维生素(B12Cobalamin) 1.36
氯化胆碱(Choline Chloride) 13.96
叶酸(Folic Acid) 2.65
肌醇(I-inositol) 18
烟酰胺(Niacinamide) 6.5
泛酸钙(D-Calcium Pantothenate) 3.5
维生素(B6Pyridoxine Hydrochloride) 0.2
核黄素(Riboflavin) 0.22
维生素(B1Thiamine Hydrochloride) 2.17
胸苷(Thymidine) 0.8
腺嘌呤(Adenine) 0.4
腐胺P(utrescine.2HCl) 0.16
丙酮酸钠(Sodium Pyruvate) 165
氯化钙(Calcium Chloride) 116.6
氯化镁(Magnesium chloride) 57.6
硫酸镁(Magnesium sulfate) 48.84
氯化钾(Potassium Chloride) 311.8
磷酸氢二钠(Sodium Phosophate Dibasic) 397.39
氯化钠(Sodium Chloride) 6999.5
磷酸二氢钠(Sodium Phosphate Monobasic.H2O) 62.5
碳酸氢钠(Sodium Bicarbonate) 2438
Hepes 3570
次黄嘌呤(Na Hypoxanthine) 4.08
酚红(Phenol Red) 8.1
硫酸铜(Cupric Sulfate.5H2O) 0.0051
硫酸亚铁(Ferrous Sulfate.7H2O) 0.834
维生素(C Ascorbic acid) 5
牛磺酸(Taurine) 30
维生素(Eα-Tocopherol acetate) 2
胆固醇(Cholesterol) 4.5
硫辛酸(Lipoic acid) 0.11
亚油酸(Linoleic acid) 0.084
硫酸锌(Zinc Sulfate.7H2O) 10.58
硝酸铁(Ferric Nitrate) 1.011
EDTA二钠(EDTA.2Na) 3.72
柠檬酸(Citric acid.H2O) 19.2
亚硒酸钠(Na2SeO3) 0.0052
乙醇胺(Ethanolamine) 1.53
巯基乙醇(β-mercaptoethnol) 6.66
嵌段式聚醚F68(Pluronic F-68) 1000
孕酮(Progesterone) 0.006
将上述组分溶解于无热源超纯水中即可配制成适合NSO细胞大规模培养及抗体生产的无蛋白培养基。
将表达抗CD25嵌合单克隆抗体的NSO细胞(从上海某医药研究有限公司获得)在本发明所述的培养基中传代适应后,在德国B.Braun公司2升生物反应器中接种,接种密度4.08×105细胞/毫升,细胞活性维持90%以上,最大比生长速率为0.92/天,最大活细胞密度为32.14×105细胞/毫升,细胞扩增近8倍,最大单抗浓度为210毫克/升。
附图1为NSO细胞在本发明无蛋白培养基批培养中的生长曲线的坐标图,附中,1为活细胞密度曲线,2为细胞活性曲线。附图2为NSO细胞在本发明无蛋白培养基批培养中的产物表达曲线的坐标图,附中,3为单抗浓度曲线。从附图的曲线可以得知,本发明适于NSO细胞大规模培养及抗体生产的无蛋白培养基能良好地支持NSO细胞的正常生长,其最高细胞密度及抗体产量均已达到或超过目前已知的商业培养基。
Claims (9)
1、一种适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,包括氨基酸、维生素、盐类、脂类、微量元素、激素、缓冲剂及葡萄糖、柠檬酸、牛磺酸、胸苷、腺嘌呤、次黄嘌呤、酚红、巯基乙醇、Pluronic F-68。
2、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的氨基酸为丙氨酸、精氨酸、天冬酰胺、天冬氨酸、半胱氨酸、胱氨酸、谷氨酸、谷胺酰胺、甘氨酸、组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、丝氨酸、苏氨酸、色氨酸、酪氨酸、缬氨酸。
3、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的维生素为生物素、维生素B12、氯化胆碱、叶酸、肌醇、烟酰胺、泛酸钙、维生素B6、核黄素、维生素B1、腐胺、维生素C、维生素E。
4、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的盐类为氯化钙、氯化镁、硫酸镁、氯化钾、磷酸氢二钠、氯化钠、磷酸二氢钠、丙酮酸钠。
5、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的脂类为胆固醇、硫辛酸、亚油酸、乙醇胺。
6、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的微量元素为硫酸铜、硫酸亚铁、硫酸锌、硝酸铁、亚硒酸钠、EDTA.2Na。
7、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的激素为孕酮。
8、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,所述的缓冲剂为碳酸氢钠和Hepes。
9、根据权利要求1所述的适于NSO细胞大规模培养及抗体生产的无蛋白培养基,其特征在于,其各种成分的含量为:mg/L
丙氨酸 80.5~104.45
精氨酸 280.5~301.2
天冬酰胺 150.5~202.5
天冬氨酸 70~106.65
半胱氨酸 55.56~67.56
胱氨酸 57~75.73
谷氨酸 800~1191
谷胺酰胺 230~292
甘氨酸 57~68.75
组氨酸 55~73.48
异亮氨酸 80.34~106.87
亮氨酸 83~111.45
赖氨酸 130~163.75
甲硫氨酸 25~32.34
苯丙氨酸 50~68.48
脯氨酸 66.45~84.50
丝氨酸 80~100
苏氨酸 70~101.05
色氨酸 15.20~19.22
酪氨酸 70~108.26
缬氨酸 80.45~99.65
葡萄糖 3500~4320
生物素 0.08~0.17
维生素B12 1.0~2.0
氯化胆碱 6.9~21
叶酸 1.4~4.2
肌醇 9~36
烟酰胺 3.3~13
泛酸钙 1.8~7
维生素 0.1~0.5
核黄素 0.11~0.46
维生素 1.1~4.4
胸苷 0.4~1.6
腺嘌呤 0.2~1
腐胺P 0.08~0.4
丙酮酸钠 165~330
氯化钙 81.6~152
氯化镁 40.3~74.8
硫酸镁 34.2~63.4
氯化钾 218~404
磷酸氢二钠 397.39
氯化钠 6999.5
磷酸二氢钠 62.5
碳酸氢钠 2438
Hepes 2000~5000
次黄嘌呤 2.04~8.16
酚红 8.1
硫酸铜 0.004~0.012
硫酸亚铁 0.62~0.96
维生素 4~10
牛磺酸 20~50
维生素 1~4
胆固醇 3.5~6.0
硫辛酸 0.08~0.20
亚油酸 0.06~0.15
硫酸锌 7.1~14.1
硝酸铁 0.85~1.21
EDTA二钠 1.0~5.2
柠檬酸 5.0~20
亚硒酸钠 0.002~0.008
乙醇胺 0~1.53
巯基乙醇 0~6.66
嵌段式聚醚F68 5000~2000
孕酮 0.003~0.012。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2008100393058A CN101418330B (zh) | 2008-06-20 | 2008-06-20 | 适于nso细胞大规模培养及抗体生产的无蛋白培养基 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2008100393058A CN101418330B (zh) | 2008-06-20 | 2008-06-20 | 适于nso细胞大规模培养及抗体生产的无蛋白培养基 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN101418330A true CN101418330A (zh) | 2009-04-29 |
CN101418330B CN101418330B (zh) | 2012-01-04 |
Family
ID=40629332
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN2008100393058A Expired - Fee Related CN101418330B (zh) | 2008-06-20 | 2008-06-20 | 适于nso细胞大规模培养及抗体生产的无蛋白培养基 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN101418330B (zh) |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104212769A (zh) * | 2014-07-14 | 2014-12-17 | 北京益生合生物科技有限公司 | 一种用于高产单抗的细胞培养基添加剂 |
CN104911143A (zh) * | 2015-06-26 | 2015-09-16 | 四川百诺吉科技有限公司 | 一种无蛋白、无水解物的无血清培养基及其制备方法 |
CN106754693A (zh) * | 2016-12-24 | 2017-05-31 | 严志海 | 一种含甘草多糖的无动物来源低蛋白培养基 |
CN107022528A (zh) * | 2017-05-18 | 2017-08-08 | 武汉博士德生物工程有限公司 | 一种用于培养杂交瘤细胞的培养基及其应用 |
CN109402040A (zh) * | 2017-08-15 | 2019-03-01 | 上海倍谙基生物科技有限公司 | 适合无血清全悬浮培养的bhk细胞及其驯化方法 |
CN109892320A (zh) * | 2019-03-20 | 2019-06-18 | 浙江省人民医院 | 一种细胞保存液及其制备方法和应用 |
CN110951670A (zh) * | 2013-03-15 | 2020-04-03 | 豪夫迈·罗氏有限公司 | 具有抗氧化剂的细胞培养组合物和多肽生产方法 |
CN111019883A (zh) * | 2019-12-24 | 2020-04-17 | 依科赛生物科技(太仓)有限公司 | Cho细胞悬浮培养的无血清无蛋白培养基及其用途 |
CN113088480A (zh) * | 2019-12-23 | 2021-07-09 | 信达生物制药(苏州)有限公司 | 一种用于cho细胞的培养基及其用途 |
CN113957042A (zh) * | 2020-07-20 | 2022-01-21 | 维他利肤(北京)生物科技有限公司 | 一种干细胞零蛋白培养基及干细胞生长因子的制备方法 |
WO2023045140A1 (zh) * | 2021-09-26 | 2023-03-30 | 上海迈泰君奥生物技术有限公司 | 一种提高宿主细胞中抗体表达量的方法 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1180080C (zh) * | 2002-01-18 | 2004-12-15 | 中国科学院上海细胞生物学研究所 | 一种改进的肝癌鼠源单克隆抗体的制备方法 |
CN100506977C (zh) * | 2007-12-28 | 2009-07-01 | 天津百若克医药生物技术有限责任公司 | 一种中国仓鼠卵巢细胞扩增培养基及其用途 |
-
2008
- 2008-06-20 CN CN2008100393058A patent/CN101418330B/zh not_active Expired - Fee Related
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110951670A (zh) * | 2013-03-15 | 2020-04-03 | 豪夫迈·罗氏有限公司 | 具有抗氧化剂的细胞培养组合物和多肽生产方法 |
CN104212769A (zh) * | 2014-07-14 | 2014-12-17 | 北京益生合生物科技有限公司 | 一种用于高产单抗的细胞培养基添加剂 |
CN104911143A (zh) * | 2015-06-26 | 2015-09-16 | 四川百诺吉科技有限公司 | 一种无蛋白、无水解物的无血清培养基及其制备方法 |
CN104911143B (zh) * | 2015-06-26 | 2019-01-22 | 四川百诺吉科技有限公司 | 一种无蛋白、无水解物的无血清培养基及其制备方法 |
CN106754693A (zh) * | 2016-12-24 | 2017-05-31 | 严志海 | 一种含甘草多糖的无动物来源低蛋白培养基 |
CN107022528A (zh) * | 2017-05-18 | 2017-08-08 | 武汉博士德生物工程有限公司 | 一种用于培养杂交瘤细胞的培养基及其应用 |
CN109402040A (zh) * | 2017-08-15 | 2019-03-01 | 上海倍谙基生物科技有限公司 | 适合无血清全悬浮培养的bhk细胞及其驯化方法 |
CN109892320A (zh) * | 2019-03-20 | 2019-06-18 | 浙江省人民医院 | 一种细胞保存液及其制备方法和应用 |
CN113088480A (zh) * | 2019-12-23 | 2021-07-09 | 信达生物制药(苏州)有限公司 | 一种用于cho细胞的培养基及其用途 |
CN113088480B (zh) * | 2019-12-23 | 2022-10-11 | 信达生物制药(苏州)有限公司 | 一种用于cho细胞的培养基及其用途 |
CN111019883A (zh) * | 2019-12-24 | 2020-04-17 | 依科赛生物科技(太仓)有限公司 | Cho细胞悬浮培养的无血清无蛋白培养基及其用途 |
CN113957042A (zh) * | 2020-07-20 | 2022-01-21 | 维他利肤(北京)生物科技有限公司 | 一种干细胞零蛋白培养基及干细胞生长因子的制备方法 |
WO2023045140A1 (zh) * | 2021-09-26 | 2023-03-30 | 上海迈泰君奥生物技术有限公司 | 一种提高宿主细胞中抗体表达量的方法 |
Also Published As
Publication number | Publication date |
---|---|
CN101418330B (zh) | 2012-01-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101418330B (zh) | 适于nso细胞大规模培养及抗体生产的无蛋白培养基 | |
CN100362098C (zh) | 适用于多种动物细胞大规模培养的无血清培养基 | |
CN101603026B (zh) | 适于动物细胞产品生产的无动物来源低蛋白培养基 | |
Yao et al. | Animal‐cell culture media: History, characteristics, and current issues | |
KR101362805B1 (ko) | 개선된 세포 배양 배지 | |
RU2644651C2 (ru) | Среда для культивирования клеток | |
CN101760442A (zh) | 适于mdck细胞大规模贴壁培养和单细胞悬浮培养的无血清培养基 | |
KR102109463B1 (ko) | 소형 펩티드를 포함하는 세포 배양 배지 | |
RU2563353C2 (ru) | Улучшенная среда для культивирования клеток | |
CN102827804B (zh) | 适用于Vero细胞微载体悬浮放大培养的培养基及方法 | |
CN107460159B (zh) | 无血清、无蛋白补料培养基及其制备方法和运用 | |
CN101864393B (zh) | 一种用于Vero细胞微载体培养的无动物来源成分无血清培养基 | |
CN101974481A (zh) | 一种用于多种肾脏组织来源细胞生长的无血清培养基 | |
CN109234223B (zh) | 低蛋白无血清细胞培养基 | |
CN1962857A (zh) | 一种哺乳动物细胞无血清培养基 | |
JP6479974B2 (ja) | 大規模エクリズマブ産生細胞培養を再現する方法 | |
CN101988047A (zh) | 一种低成本的昆虫细胞无血清培养基 | |
CN102317440A (zh) | 改进的培养基添加剂及其应用方法 | |
CN104328158B (zh) | 适于动物细胞表达产品大规模生产的化学成分明确培养基 | |
CN105462912A (zh) | 适用于二倍体细胞培养的无蛋白无血清培养基及应用 | |
CN106399224A (zh) | 无血清无蛋白细胞培养基 | |
Burgener et al. | Medium development | |
IE921020A1 (en) | Serum-free medium for culturing mammalian cells | |
CN106754634A (zh) | 一种无血清培养基及其制备方法 | |
Chen et al. | A low-cost chemically defined protein free medium for a recombinant CHO cell line producing prothrombin |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120104 |