CN1551918B - 基因重组伊卡因及其制备方法 - Google Patents
基因重组伊卡因及其制备方法 Download PDFInfo
- Publication number
- CN1551918B CN1551918B CN028175026A CN02817502A CN1551918B CN 1551918 B CN1551918 B CN 1551918B CN 028175026 A CN028175026 A CN 028175026A CN 02817502 A CN02817502 A CN 02817502A CN 1551918 B CN1551918 B CN 1551918B
- Authority
- CN
- China
- Prior art keywords
- kayin
- gene
- gene recombination
- preparation
- cell
- 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.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims description 24
- 108010085662 ecarin Proteins 0.000 title abstract description 9
- 230000008569 process Effects 0.000 title description 5
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 57
- 239000013604 expression vector Substances 0.000 claims abstract description 14
- 239000012228 culture supernatant Substances 0.000 claims abstract description 12
- 244000005700 microbiome Species 0.000 claims abstract description 9
- 210000004027 cell Anatomy 0.000 claims description 48
- 238000005215 recombination Methods 0.000 claims description 25
- 230000006798 recombination Effects 0.000 claims description 23
- 238000002360 preparation method Methods 0.000 claims description 21
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 19
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 13
- 239000012634 fragment Substances 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 8
- 238000005277 cation exchange chromatography Methods 0.000 claims description 7
- 238000005227 gel permeation chromatography Methods 0.000 claims description 5
- 239000002773 nucleotide Substances 0.000 claims description 5
- 125000003729 nucleotide group Chemical group 0.000 claims description 5
- 102000007469 Actins Human genes 0.000 claims description 4
- 108010085238 Actins Proteins 0.000 claims description 4
- 241000271566 Aves Species 0.000 claims description 4
- 230000014509 gene expression Effects 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 3
- 241000701022 Cytomegalovirus Species 0.000 claims description 2
- 108060003951 Immunoglobulin Proteins 0.000 claims description 2
- 241001529936 Murinae Species 0.000 claims description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 2
- 108700007696 Tetrahydrofolate Dehydrogenase Proteins 0.000 claims description 2
- IXKSXJFAGXLQOQ-XISFHERQSA-N WHWLQLKPGQPMY Chemical group C([C@@H](C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CNC=N1 IXKSXJFAGXLQOQ-XISFHERQSA-N 0.000 claims description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 claims description 2
- 102000004419 dihydrofolate reductase Human genes 0.000 claims description 2
- 102000018358 immunoglobulin Human genes 0.000 claims description 2
- 201000000050 myeloid neoplasm Diseases 0.000 claims description 2
- 108020001775 protein parts Proteins 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims description 2
- 238000011144 upstream manufacturing Methods 0.000 claims description 2
- 101150084750 1 gene Proteins 0.000 claims 3
- 101150028074 2 gene Proteins 0.000 claims 3
- 230000004544 DNA amplification Effects 0.000 claims 3
- 238000009825 accumulation Methods 0.000 claims 1
- 239000012535 impurity Substances 0.000 claims 1
- 108010094028 Prothrombin Proteins 0.000 abstract description 11
- 102100027378 Prothrombin Human genes 0.000 abstract description 11
- 229940039716 prothrombin Drugs 0.000 abstract description 11
- 102000004169 proteins and genes Human genes 0.000 abstract description 10
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 238000012258 culturing Methods 0.000 abstract description 5
- 210000004102 animal cell Anatomy 0.000 abstract description 2
- 238000010353 genetic engineering Methods 0.000 abstract description 2
- 108010064129 Thrombogen Proteins 0.000 description 20
- 230000008521 reorganization Effects 0.000 description 20
- 230000000694 effects Effects 0.000 description 19
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 15
- 108091008146 restriction endonucleases Proteins 0.000 description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 101150074155 DHFR gene Proteins 0.000 description 13
- QYLJIYOGHRGUIH-CIUDSAMLSA-N Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CCCNC(N)=N QYLJIYOGHRGUIH-CIUDSAMLSA-N 0.000 description 12
- 108010011227 meizothrombin Proteins 0.000 description 12
- 238000013016 damping Methods 0.000 description 11
- 239000013613 expression plasmid Substances 0.000 description 11
- 239000012530 fluid Substances 0.000 description 11
- 230000029087 digestion Effects 0.000 description 10
- 239000002299 complementary DNA Substances 0.000 description 9
- 238000000502 dialysis Methods 0.000 description 9
- 108020004707 nucleic acids Proteins 0.000 description 9
- 102000039446 nucleic acids Human genes 0.000 description 9
- 150000007523 nucleic acids Chemical class 0.000 description 9
- 108010071286 prethrombins Proteins 0.000 description 9
- 235000018102 proteins Nutrition 0.000 description 9
- 238000005336 cracking Methods 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 8
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 8
- 102000012410 DNA Ligases Human genes 0.000 description 7
- 108010061982 DNA Ligases Proteins 0.000 description 7
- 230000004913 activation Effects 0.000 description 7
- 229920001184 polypeptide Polymers 0.000 description 7
- 102000004196 processed proteins & peptides Human genes 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- 101100230376 Acetivibrio thermocellus (strain ATCC 27405 / DSM 1237 / JCM 9322 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372) celI gene Proteins 0.000 description 6
- 241000699802 Cricetulus griseus Species 0.000 description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 238000004587 chromatography analysis Methods 0.000 description 6
- 210000001672 ovary Anatomy 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 239000003998 snake venom Substances 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 230000003213 activating effect Effects 0.000 description 5
- 235000001014 amino acid Nutrition 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 230000026731 phosphorylation Effects 0.000 description 5
- 238000006366 phosphorylation reaction Methods 0.000 description 5
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 4
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000002523 gelfiltration Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000012679 serum free medium Substances 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- XNSKSTRGQIPTSE-ACZMJKKPSA-N Arg-Thr Chemical compound C[C@@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O XNSKSTRGQIPTSE-ACZMJKKPSA-N 0.000 description 3
- 101000962067 Gallus gallus Neuroblastoma suppressor of tumorigenicity 1 Proteins 0.000 description 3
- 239000006180 TBST buffer Substances 0.000 description 3
- 108090000190 Thrombin Proteins 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 244000309466 calf Species 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000004520 electroporation Methods 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000002161 passivation Methods 0.000 description 3
- -1 polyoxyethylene Polymers 0.000 description 3
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 235000017557 sodium bicarbonate Nutrition 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 229960004072 thrombin Drugs 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- YDMBNDUHUNWWRP-VJBWXMMDSA-N (2s)-1-[(2r)-2-amino-3-phenylpropanoyl]-n-[(2s)-5-(diaminomethylideneamino)-1-(4-nitroanilino)-1-oxopentan-2-yl]piperidine-2-carboxamide Chemical compound C([C@@H](N)C(=O)N1[C@@H](CCCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NC=1C=CC(=CC=1)[N+]([O-])=O)C1=CC=CC=C1 YDMBNDUHUNWWRP-VJBWXMMDSA-N 0.000 description 2
- PJWWRFATQTVXHA-UHFFFAOYSA-N Cyclohexylaminopropanesulfonic acid Chemical compound OS(=O)(=O)CCCNC1CCCCC1 PJWWRFATQTVXHA-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 108020000311 Glutamate Synthase Proteins 0.000 description 2
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 2
- 108010039286 S 2238 Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 108010022999 Serine Proteases Proteins 0.000 description 2
- 102000012479 Serine Proteases Human genes 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 238000001042 affinity chromatography Methods 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 230000009849 deactivation Effects 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000002489 hematologic effect Effects 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229920001993 poloxamer 188 Polymers 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- 239000002435 venom Substances 0.000 description 2
- 231100000611 venom Toxicity 0.000 description 2
- 210000001048 venom Anatomy 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 101710154825 Aminoglycoside 3'-phosphotransferase Proteins 0.000 description 1
- 102000004411 Antithrombin III Human genes 0.000 description 1
- 108090000935 Antithrombin III Proteins 0.000 description 1
- 241000239290 Araneae Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 241000282552 Chlorocebus aethiops Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102100024746 Dihydrofolate reductase Human genes 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000122860 Echis carinatus Species 0.000 description 1
- KGWDUNBJIMUFAP-KVVVOXFISA-N Ethanolamine Oleate Chemical compound NCCO.CCCCCCCC\C=C/CCCCCCCC(O)=O KGWDUNBJIMUFAP-KVVVOXFISA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102100024319 Intestinal-type alkaline phosphatase Human genes 0.000 description 1
- 101710184243 Intestinal-type alkaline phosphatase Proteins 0.000 description 1
- 108010025815 Kanamycin Kinase Proteins 0.000 description 1
- 108010006035 Metalloproteases Proteins 0.000 description 1
- 102000005741 Metalloproteases Human genes 0.000 description 1
- 125000000729 N-terminal amino-acid group Chemical group 0.000 description 1
- 101710138657 Neurotoxin Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 108090000040 Russellysin Proteins 0.000 description 1
- 101100221606 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) COS7 gene Proteins 0.000 description 1
- 241000270295 Serpentes Species 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 102000002070 Transferrins Human genes 0.000 description 1
- 108010015865 Transferrins Proteins 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229930003448 Vitamin K Natural products 0.000 description 1
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 102000006646 aminoglycoside phosphotransferase Human genes 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229960005348 antithrombin iii Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 239000003659 bee venom Substances 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 230000030609 dephosphorylation Effects 0.000 description 1
- 238000006209 dephosphorylation reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 108020001096 dihydrofolate reductase Proteins 0.000 description 1
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 230000001723 fibrinogenic effect Effects 0.000 description 1
- 210000002196 fr. b Anatomy 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 108010039551 hemorrhagic proteinase IV Proteins 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002505 iron Chemical class 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 238000001638 lipofection Methods 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 239000002581 neurotoxin Substances 0.000 description 1
- 231100000618 neurotoxin Toxicity 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 238000007500 overflow downdraw method Methods 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000012716 precipitator Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000000734 protein sequencing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 239000002795 scorpion venom Substances 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012882 sequential analysis Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 229960001471 sodium selenite Drugs 0.000 description 1
- 235000015921 sodium selenite Nutrition 0.000 description 1
- 239000011781 sodium selenite Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 239000002821 viper venom Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6402—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals
- C12N9/6418—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals from snakes
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims (11)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP206918/2001 | 2001-07-06 | ||
JP2001206918 | 2001-07-06 | ||
PCT/JP2002/006770 WO2003004647A1 (fr) | 2001-07-06 | 2002-07-04 | Ecarine genetiquement modifiee et procede d'elaboration |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1551918A CN1551918A (zh) | 2004-12-01 |
CN1551918B true CN1551918B (zh) | 2011-04-20 |
Family
ID=19042978
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN028175026A Expired - Fee Related CN1551918B (zh) | 2001-07-06 | 2002-07-04 | 基因重组伊卡因及其制备方法 |
Country Status (7)
Country | Link |
---|---|
US (2) | US20050164365A1 (zh) |
EP (1) | EP1405912B8 (zh) |
JP (1) | JP4150814B2 (zh) |
KR (1) | KR101054735B1 (zh) |
CN (1) | CN1551918B (zh) |
CA (1) | CA2452767C (zh) |
WO (1) | WO2003004647A1 (zh) |
Families Citing this family (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB0224986D0 (en) | 2002-10-28 | 2002-12-04 | Smith & Nephew | Apparatus |
GB0324044D0 (en) | 2003-10-14 | 2003-11-19 | Astrazeneca Ab | Protein |
US8529548B2 (en) | 2004-04-27 | 2013-09-10 | Smith & Nephew Plc | Wound treatment apparatus and method |
GB0409444D0 (en) * | 2004-04-28 | 2004-06-02 | Smith & Nephew | Apparatus |
GB0409446D0 (en) | 2004-04-28 | 2004-06-02 | Smith & Nephew | Apparatus |
RU2415934C2 (ru) | 2005-04-13 | 2011-04-10 | Астразенека Аб | Клетка-хозяин, содержащая вектор для продуцирования белков, требующих гамма-карбоксилирования |
JP2009528843A (ja) * | 2006-03-06 | 2009-08-13 | ヒューマジーン・インコーポレイテッド | 組換えヒトトロンビンおよびフィブリノゲンの調製法 |
US8206967B2 (en) | 2007-07-06 | 2012-06-26 | Medimmune Limited | Method for production of recombinant human thrombin |
CA2716738A1 (en) * | 2008-04-07 | 2009-10-15 | Zymogenetics, Inc. | Thrombin activator compositions and methods of making and using the same |
CN103026223B (zh) * | 2010-06-18 | 2015-02-25 | 株式会社大赛璐 | 光学异构体用分离剂 |
US10597440B2 (en) | 2014-01-29 | 2020-03-24 | Km Biologics Co., Ltd. | Anti-transthyretin human antibody |
KR102538555B1 (ko) | 2014-01-29 | 2023-05-30 | 케이엠 바이올로직스 가부시키가이샤 | 항-트랜스티레틴 인간화 항체 |
JPWO2020085457A1 (ja) | 2018-10-25 | 2021-09-30 | Kmバイオロジクス株式会社 | 改変CMV gBタンパク質及びこれを含むCMVワクチン |
WO2020121983A1 (ja) | 2018-12-10 | 2020-06-18 | Kmバイオロジクス株式会社 | サイトメガロウイルスの先天性感染を予防又は治療するためのワクチン |
WO2020175660A1 (ja) | 2019-02-28 | 2020-09-03 | Kmバイオロジクス株式会社 | Rsv f/gキメラワクチン |
CA3186423C (en) | 2020-06-09 | 2024-03-19 | Km Biologics Co., Ltd. | Fusion protein of pentamer and gb of cytomegalovirus, and vaccine containing said fusion protein |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA1340772C (en) * | 1987-12-30 | 1999-09-28 | Patricia Tekamp-Olson | Expression and secretion of heterologous protiens in yeast employing truncated alpha-factor leader sequences |
CA2078721A1 (en) * | 1991-09-24 | 1993-03-25 | Hiroshi Yonemura | Process for preparing human coagulation factor viii protein complex |
AU702379B2 (en) * | 1996-06-04 | 1999-02-18 | Juridical Foundation The Chemo-Sero-Therapeutic Research Institute | Novel feline cytokine protein |
US6413737B1 (en) * | 1999-07-09 | 2002-07-02 | Cohesion Technologies, Inc. | Ecarin prothrombin protease and methods |
-
2002
- 2002-07-04 KR KR1020047000117A patent/KR101054735B1/ko active IP Right Grant
- 2002-07-04 WO PCT/JP2002/006770 patent/WO2003004647A1/ja active Application Filing
- 2002-07-04 CN CN028175026A patent/CN1551918B/zh not_active Expired - Fee Related
- 2002-07-04 JP JP2003510805A patent/JP4150814B2/ja not_active Expired - Lifetime
- 2002-07-04 US US10/482,925 patent/US20050164365A1/en not_active Abandoned
- 2002-07-04 CA CA2452767A patent/CA2452767C/en not_active Expired - Fee Related
- 2002-07-04 EP EP02745822A patent/EP1405912B8/en not_active Expired - Lifetime
-
2007
- 2007-12-05 US US11/950,638 patent/US20080153132A1/en not_active Abandoned
Non-Patent Citations (2)
Title |
---|
Giulia Russo 等.Stable expression and purification of a secreted humanrecombinant prethrombin-2 and its activation to thrombin.Protein Expression and Purification 10.1997,(10),214-225. |
Giulia Russo等.Stable expression and purification of a secreted humanrecombinant prethrombin-2 and its activation to thrombin.Protein Expression and Purification 10.1997,(10),214-225. * |
Also Published As
Publication number | Publication date |
---|---|
CA2452767A1 (en) | 2003-01-16 |
WO2003004647A1 (fr) | 2003-01-16 |
CN1551918A (zh) | 2004-12-01 |
EP1405912A4 (en) | 2005-01-05 |
KR101054735B1 (ko) | 2011-08-05 |
EP1405912B8 (en) | 2012-10-24 |
CA2452767C (en) | 2013-03-19 |
US20080153132A1 (en) | 2008-06-26 |
KR20040018427A (ko) | 2004-03-03 |
EP1405912B1 (en) | 2012-09-05 |
JPWO2003004647A1 (ja) | 2004-10-28 |
EP1405912A1 (en) | 2004-04-07 |
JP4150814B2 (ja) | 2008-09-17 |
US20050164365A1 (en) | 2005-07-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1551918B (zh) | 基因重组伊卡因及其制备方法 | |
James et al. | Transport of proteins into chloroplasts: import and maturation of precursors to the 33-, 23-, and 16-kDa proteins of the photosynthetic oxygen-evolving complex | |
Grinnell et al. | Trans–Activated Expression of Fully Gamma–Carboxylated Recombinant Human Protein C, an Antithrombotic Factor | |
EP0612350A1 (en) | Ubiquitin-specific proteases | |
CA2066174A1 (en) | Process for the enzymatic preparation of basic fibroblast growth factor | |
USRE37958E1 (en) | DNA sequence coding for protein C | |
EP1405910B1 (en) | Process for producing human thrombin by gene modification technique | |
CN103773780B (zh) | 巴曲酶的突变核苷酸序列、突变的α因子分泌信号序列和使用其制备巴曲酶的方法 | |
US4923818A (en) | DNA clone of human type IV collagenase | |
US8017750B2 (en) | Haemocoagulase | |
CA1339874C (en) | Dna clone of human type iv collagenase | |
AU2008202374B2 (en) | Genetically modified ecarin and process for producing the same | |
KR0180103B1 (ko) | 바실러스 서브틸리스 속 균주 유래의 혈전용해효소 | |
CN100424172C (zh) | 一种定向突变的基因工程巴曲酶及用途 | |
Hatsuzawa et al. | Purification of mouse Ren 2 prorenin produced in Chinese hamster ovary cells | |
JP2009502118A (ja) | 敗血症治療のための組み換え活性化ヒトタンパク質cを生成する方法 | |
Macgillivray et al. | 6 Molecular biology of factor X | |
USRE38981E1 (en) | DNA sequence coding for protein C | |
AU2008202376B2 (en) | Process for producing human thrombin by gene modification technique | |
KR20090116669A (ko) | 배트록소빈의 변이 뉴클레오타이드 서열, α-인자 분비 시그널 서열 변이체 및 이를 이용한 재조합 단백질의 제조방법및 이를 이용한 배트록소빈의 제조방법 | |
Lee et al. | Studies on the Development of a Thrombolytic Agent from Korean Snake Venom I. Purification of a Protease from the Venom of A. bromhoffi brevicaudus |
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 | ||
CP03 | Change of name, title or address |
Address after: Xiongben County, Xiongben, Japan Patentee after: THE CHEMO-SERO-THERAPEUTIC Research Institute Address before: Kumamoto Prefecture, Japan Patentee before: JURIDICAL FOUNDATION THE CHEMO-SERO-THERAPEUTIC Research Institute |
|
CP03 | Change of name, title or address | ||
TR01 | Transfer of patent right |
Effective date of registration: 20200309 Address after: Kumamoto, Japan Patentee after: THE CHEMO-SERO-THERAPEUTIC Research Institute Address before: Xiongben County, Xiongben, Japan Patentee before: THE CHEMO-SERO-THERAPEUTIC Research Institute |
|
TR01 | Transfer of patent right | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110420 Termination date: 20200704 |
|
CF01 | Termination of patent right due to non-payment of annual fee |