NO331376B1 - DNA-molekyl kodende for fusjonsproteiner der den N-terminale delen er et hirudinderivat for fremstilling av rekombinante proteiner via sekresjon fra gjaer, samt multikopivektor, plasmid, vertscelle og fremgangsmater for fermentering og fremstilling - Google Patents
DNA-molekyl kodende for fusjonsproteiner der den N-terminale delen er et hirudinderivat for fremstilling av rekombinante proteiner via sekresjon fra gjaer, samt multikopivektor, plasmid, vertscelle og fremgangsmater for fermentering og fremstilling Download PDFInfo
- Publication number
- NO331376B1 NO331376B1 NO20033672A NO20033672A NO331376B1 NO 331376 B1 NO331376 B1 NO 331376B1 NO 20033672 A NO20033672 A NO 20033672A NO 20033672 A NO20033672 A NO 20033672A NO 331376 B1 NO331376 B1 NO 331376B1
- Authority
- NO
- Norway
- Prior art keywords
- hirudin
- protein
- dna
- fusion protein
- plasmid
- Prior art date
Links
- WQPDUTSPKFMPDP-OUMQNGNKSA-N hirudin Chemical class C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(OS(O)(=O)=O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]1NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]2CSSC[C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@H](C(NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N2)=O)CSSC1)C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=2C=CC(O)=CC=2)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)CSSC1)C(C)C)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 WQPDUTSPKFMPDP-OUMQNGNKSA-N 0.000 title claims abstract description 46
- 108020004414 DNA Proteins 0.000 title claims abstract description 35
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 20
- 108020001507 fusion proteins Proteins 0.000 title claims description 36
- 102000037865 fusion proteins Human genes 0.000 title claims description 36
- 239000013612 plasmid Substances 0.000 title claims description 26
- 239000013598 vector Substances 0.000 title claims description 21
- 102000053602 DNA Human genes 0.000 title claims description 14
- 238000000855 fermentation Methods 0.000 title claims description 11
- 230000004151 fermentation Effects 0.000 title claims description 11
- 238000004519 manufacturing process Methods 0.000 title description 10
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 title description 4
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 title description 4
- 230000028327 secretion Effects 0.000 title description 3
- 238000002360 preparation method Methods 0.000 title 1
- 229940006607 hirudin Drugs 0.000 claims abstract description 41
- 108010007267 Hirudins Proteins 0.000 claims abstract description 40
- 102000007625 Hirudins Human genes 0.000 claims abstract description 39
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 27
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 25
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 17
- 108020004705 Codon Proteins 0.000 claims abstract description 14
- 150000001413 amino acids Chemical class 0.000 claims abstract description 14
- 239000000126 substance Substances 0.000 claims abstract description 9
- 108010076504 Protein Sorting Signals Proteins 0.000 claims abstract description 3
- 101710188306 Protein Y Proteins 0.000 claims abstract description 3
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 29
- 210000004027 cell Anatomy 0.000 claims description 22
- 238000000034 method Methods 0.000 claims description 21
- 229940125396 insulin Drugs 0.000 claims description 17
- 108010013359 miniproinsulin Proteins 0.000 claims description 16
- 239000006228 supernatant Substances 0.000 claims description 16
- 102000004877 Insulin Human genes 0.000 claims description 12
- 108090001061 Insulin Proteins 0.000 claims description 12
- 241000235058 Komagataella pastoris Species 0.000 claims description 7
- 108090000631 Trypsin Proteins 0.000 claims description 7
- 102000004142 Trypsin Human genes 0.000 claims description 7
- 239000012588 trypsin Substances 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 4
- 238000001556 precipitation Methods 0.000 claims description 4
- 102000003670 Carboxypeptidase B Human genes 0.000 claims description 3
- 108090000087 Carboxypeptidase B Proteins 0.000 claims description 3
- 238000004113 cell culture Methods 0.000 claims description 3
- 210000000349 chromosome Anatomy 0.000 claims description 3
- 239000012228 culture supernatant Substances 0.000 claims description 3
- 238000001471 micro-filtration Methods 0.000 claims description 3
- 241001138401 Kluyveromyces lactis Species 0.000 claims description 2
- 239000012737 fresh medium Substances 0.000 claims description 2
- 238000004191 hydrophobic interaction chromatography Methods 0.000 claims description 2
- 238000004255 ion exchange chromatography Methods 0.000 claims description 2
- 239000011541 reaction mixture Substances 0.000 claims description 2
- 241000125500 Hedypnois rhagadioloides Species 0.000 claims 1
- 235000018102 proteins Nutrition 0.000 description 17
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 16
- 239000000047 product Substances 0.000 description 10
- 102000001708 Protein Isoforms Human genes 0.000 description 8
- 108010029485 Protein Isoforms Proteins 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 229940024606 amino acid Drugs 0.000 description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000012634 fragment Substances 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 108010002230 lepirudin Proteins 0.000 description 5
- FIBJDTSHOUXTKV-BRHMIFOHSA-N lepirudin Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](Cc1ccccc1)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](Cc1c[nH]cn1)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCCN)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@@H]1CSSC[C@@H]2NC(=O)[C@@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CSSC[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)CNC2=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N1)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](Cc2ccc(O)cc2)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)[C@@H](C)O)[C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N1)C(C)C)C(C)C)[C@@H](C)O)[C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O FIBJDTSHOUXTKV-BRHMIFOHSA-N 0.000 description 5
- 229940030915 refludan Drugs 0.000 description 5
- 238000010276 construction Methods 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 102100034044 All-trans-retinol dehydrogenase [NAD(+)] ADH1B Human genes 0.000 description 2
- 101710193111 All-trans-retinol dehydrogenase [NAD(+)] ADH4 Proteins 0.000 description 2
- 101001002617 Bos taurus Interleukin-2 Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 108010076181 Proinsulin Proteins 0.000 description 2
- 238000012300 Sequence Analysis Methods 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- IXKSXJFAGXLQOQ-XISFHERQSA-N WHWLQLKPGQPMY Chemical compound 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 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000005277 cation exchange chromatography Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 238000007169 ligase reaction Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000004007 reversed phase HPLC Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- QIJRTFXNRTXDIP-UHFFFAOYSA-N (1-carboxy-2-sulfanylethyl)azanium;chloride;hydrate Chemical compound O.Cl.SCC(N)C(O)=O QIJRTFXNRTXDIP-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 101100519159 Arabidopsis thaliana PCR3 gene Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- 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 description 1
- XCLCVBYNGXEVDU-WHFBIAKZSA-N Gly-Asn-Ser Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O XCLCVBYNGXEVDU-WHFBIAKZSA-N 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 1
- 241000545744 Hirudinea Species 0.000 description 1
- 241000237903 Hirudo Species 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 101500021084 Locusta migratoria 5 kDa peptide Proteins 0.000 description 1
- 108010074338 Lymphokines Proteins 0.000 description 1
- 102000008072 Lymphokines Human genes 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 description 1
- 241000320412 Ogataea angusta Species 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 101710118538 Protease Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 101000712605 Theromyzon tessulatum Theromin Proteins 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 229940122388 Thrombin inhibitor Drugs 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 108010084455 Zeocin Proteins 0.000 description 1
- 229910007426 ZnC2 Inorganic materials 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000010352 biotechnological method Methods 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000012993 chemical processing Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 229960001305 cysteine hydrochloride Drugs 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000009655 industrial fermentation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 229940047122 interleukins Drugs 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000013386 optimize process Methods 0.000 description 1
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 1
- 238000013492 plasmid preparation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 239000003868 thrombin inhibitor Substances 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/815—Protease inhibitors from leeches, e.g. hirudin, eglin
-
- 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
- C07K14/575—Hormones
- C07K14/62—Insulins
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE10108211A DE10108211A1 (de) | 2001-02-20 | 2001-02-20 | Verwendung von Fusionsproteinen, deren N-terminaler Anteil aus einem Hirudinderivat besteht, zur Herstellung rekombinanter Proteine über Sekretion durch Hefen |
| PCT/EP2002/001308 WO2002070722A1 (en) | 2001-02-20 | 2002-02-08 | Use of fusion proteins whose n-terminal part is a hirudin derivative for the production of recombinant proteins via secretion by yeasts |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| NO20033672D0 NO20033672D0 (no) | 2003-08-19 |
| NO20033672L NO20033672L (no) | 2003-10-17 |
| NO331376B1 true NO331376B1 (no) | 2011-12-12 |
Family
ID=7674912
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| NO20033672A NO331376B1 (no) | 2001-02-20 | 2003-08-19 | DNA-molekyl kodende for fusjonsproteiner der den N-terminale delen er et hirudinderivat for fremstilling av rekombinante proteiner via sekresjon fra gjaer, samt multikopivektor, plasmid, vertscelle og fremgangsmater for fermentering og fremstilling |
Country Status (22)
| Country | Link |
|---|---|
| US (1) | US20030176673A1 (el) |
| EP (1) | EP1364032B1 (el) |
| JP (1) | JP4199543B2 (el) |
| KR (1) | KR100858833B1 (el) |
| CN (1) | CN1279171C (el) |
| AT (1) | ATE342367T1 (el) |
| AU (1) | AU2002250903B2 (el) |
| BR (1) | BR0207379A (el) |
| CA (1) | CA2439042C (el) |
| CY (1) | CY1106274T1 (el) |
| DE (2) | DE10108211A1 (el) |
| DK (1) | DK1364032T3 (el) |
| ES (1) | ES2272696T3 (el) |
| HK (1) | HK1065068A1 (el) |
| IL (2) | IL157416A0 (el) |
| MX (1) | MXPA03007116A (el) |
| NO (1) | NO331376B1 (el) |
| NZ (1) | NZ527642A (el) |
| PE (1) | PE20020857A1 (el) |
| PT (1) | PT1364032E (el) |
| WO (1) | WO2002070722A1 (el) |
| ZA (1) | ZA200305868B (el) |
Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7638618B2 (en) * | 2001-02-20 | 2009-12-29 | Sanofi-Aventis Deutschland Gmbh | Nucleic acids encoding a hirudin and pro-insulin as superscretable peptides and for parallel improvement of the exported forms of one or more polypeptides of interest |
| DE102006031955A1 (de) | 2006-07-11 | 2008-01-17 | Sanofi-Aventis Deutschland Gmbh | Verfahren zur Herstellung von Insulinanaloga mit dibasischem B-Kettenende |
| DE102006031962A1 (de) | 2006-07-11 | 2008-01-17 | Sanofi-Aventis Deutschland Gmbh | Amidiertes Insulin Glargin |
| DE102008003566A1 (de) | 2008-01-09 | 2009-07-16 | Sanofi-Aventis Deutschland Gmbh | Neue Insulinderivate mit extrem verzögertem Zeit-/ Wirkungsprofil |
| RU2524423C2 (ru) | 2008-01-09 | 2014-07-27 | Санофи-Авентис Дойчланд Гмбх | Новые производные инсулина с чрезвычайно замедленным профилем время/действие |
| DE102008025007A1 (de) | 2008-05-24 | 2009-11-26 | Sanofi-Aventis Deutschland Gmbh | Neue Insulinderivate mit extrem verzögertem Zeit-/ Wirkungsprofil |
| WO2009087082A2 (de) | 2008-01-09 | 2009-07-16 | Sanofi-Aventis Deutschland Gmbh | Neue insulinderivate mit extrem verzögertem zeit- / wirkungsprofil |
| RS59913B1 (sr) | 2008-10-17 | 2020-03-31 | Sanofi Aventis Deutschland | Kombinacija insulina i glp-1-agonista |
| KR101972301B1 (ko) | 2009-11-13 | 2019-04-25 | 사노피-아벤티스 도이칠란트 게엠베하 | Ｇｌｐ－１ 작용제, 인슐린 및 메티오닌을 포함하는 약제학적 조성물 |
| BR112012011403B8 (pt) | 2009-11-13 | 2021-05-25 | Sanofi Aventis Deutschland | composição farmacêutica líquida compreendendo um agonista glp-1 e metionina e uso da mesma |
| CA2809321C (en) | 2010-08-30 | 2018-08-14 | Sanofi-Aventis Deutschland Gmbh | Use of ave0010 for the manufacture of a medicament for the treatment of diabetes mellitus type 2 |
| US9821032B2 (en) | 2011-05-13 | 2017-11-21 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical combination for improving glycemic control as add-on therapy to basal insulin |
| HUE027989T2 (en) | 2011-08-29 | 2016-11-28 | Sanofi Aventis Deutschland | A pharmaceutical composition for use in glycemic control in patients with type 2 diabetes |
| AR087744A1 (es) | 2011-09-01 | 2014-04-16 | Sanofi Aventis Deutschland | Composicion farmaceutica para uso en el tratamiento de una enfermedad neurodegenerativa |
| SI3229828T1 (sl) | 2014-12-12 | 2023-06-30 | Sanofi-Aventis Deutschland Gmbh | Formulacija s fiksnim razmerjem inzulin glargin/liksisenatid |
| TWI748945B (zh) | 2015-03-13 | 2021-12-11 | 德商賽諾菲阿凡提斯德意志有限公司 | 第２型糖尿病病患治療 |
| TW201705975A (zh) | 2015-03-18 | 2017-02-16 | 賽諾菲阿凡提斯德意志有限公司 | 第２型糖尿病病患之治療 |
Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0158564B1 (fr) * | 1984-03-27 | 1992-07-15 | Transgene S.A. | Vecteurs d'expression de l'hirudine, cellules transformées et procédé de préparation de l'hirudine |
| US5705355A (en) * | 1984-03-27 | 1998-01-06 | Transgene, S.A. | Hirudin, pharmaceutical compositions comprising it and their use |
| DE3738541A1 (de) * | 1987-11-13 | 1989-05-24 | Hoechst Ag | Verfahren zur isolierung und reinigung von hirudin |
| US6875589B1 (en) * | 1988-06-23 | 2005-04-05 | Hoechst Aktiengesellschaft | Mini-proinsulin, its preparation and use |
| GB8927722D0 (en) * | 1989-12-07 | 1990-02-07 | British Bio Technology | Proteins and nucleic acids |
| DE4009268A1 (de) * | 1990-03-22 | 1991-09-26 | Consortium Elektrochem Ind | Sekretion von hirudinderivaten |
| DK0511393T3 (da) * | 1990-11-08 | 1996-12-09 | Japan Energy Corp | Hirudin-mutant, fremstilling af denne, antikoagulant, sekretorisk vektor, mikroorganisme transformeret af vektoren og fremstilling af produkt ud fra mikroorganismen |
| FI92601C (fi) * | 1992-03-11 | 1994-12-12 | Marja Makarow | Menetelmä hyötyproteiinien erittämiseksi hiivoista |
| DE19543737A1 (de) * | 1995-11-24 | 1997-05-28 | Hoechst Ag | Verfahren zur Ultrafiltration von Peptide oder Proteine enthaltender biologischer Matrices |
| DE19544233A1 (de) * | 1995-11-28 | 1997-06-05 | Hoechst Ag | Verfahren zur Nutzung des Hefe-ADH II-Promotorsystems zur biotechnologischen Produktion heterologer Proteine in hohen Ausbeuten |
| DK0889949T3 (da) * | 1996-03-13 | 2003-09-22 | Delta Biotechnology Ltd | Fermenteringskontrol |
| DE10033195A1 (de) * | 2000-07-07 | 2002-03-21 | Aventis Pharma Gmbh | Bifunktionale Fusionsproteine aus Hirudin und TAP |
| US7202059B2 (en) * | 2001-02-20 | 2007-04-10 | Sanofi-Aventis Deutschland Gmbh | Fusion proteins capable of being secreted into a fermentation medium |
-
2001
- 2001-02-20 DE DE10108211A patent/DE10108211A1/de not_active Withdrawn
-
2002
- 2002-02-06 PE PE2002000082A patent/PE20020857A1/es not_active Application Discontinuation
- 2002-02-08 JP JP2002570745A patent/JP4199543B2/ja not_active Expired - Fee Related
- 2002-02-08 DE DE60215315T patent/DE60215315T2/de not_active Expired - Lifetime
- 2002-02-08 ES ES02719782T patent/ES2272696T3/es not_active Expired - Lifetime
- 2002-02-08 IL IL15741602A patent/IL157416A0/xx unknown
- 2002-02-08 WO PCT/EP2002/001308 patent/WO2002070722A1/en active IP Right Grant
- 2002-02-08 EP EP02719782A patent/EP1364032B1/en not_active Expired - Lifetime
- 2002-02-08 BR BR0207379-0A patent/BR0207379A/pt not_active Application Discontinuation
- 2002-02-08 CN CNB028068491A patent/CN1279171C/zh not_active Expired - Fee Related
- 2002-02-08 DK DK02719782T patent/DK1364032T3/da active
- 2002-02-08 KR KR1020037010932A patent/KR100858833B1/ko not_active IP Right Cessation
- 2002-02-08 MX MXPA03007116A patent/MXPA03007116A/es active IP Right Grant
- 2002-02-08 PT PT02719782T patent/PT1364032E/pt unknown
- 2002-02-08 CA CA2439042A patent/CA2439042C/en not_active Expired - Fee Related
- 2002-02-08 NZ NZ527642A patent/NZ527642A/en not_active IP Right Cessation
- 2002-02-08 AT AT02719782T patent/ATE342367T1/de active
- 2002-02-08 AU AU2002250903A patent/AU2002250903B2/en not_active Ceased
- 2002-02-19 US US10/076,632 patent/US20030176673A1/en not_active Abandoned
-
2003
- 2003-07-30 ZA ZA2003/05868A patent/ZA200305868B/en unknown
- 2003-08-14 IL IL157416A patent/IL157416A/en not_active IP Right Cessation
- 2003-08-19 NO NO20033672A patent/NO331376B1/no not_active IP Right Cessation
-
2004
- 2004-10-14 HK HK04107931A patent/HK1065068A1/xx not_active IP Right Cessation
-
2006
- 2006-11-30 CY CY20061101726T patent/CY1106274T1/el unknown
Also Published As
| Publication number | Publication date |
|---|---|
| US20030176673A1 (en) | 2003-09-18 |
| PE20020857A1 (es) | 2002-11-04 |
| DE10108211A1 (de) | 2002-08-22 |
| BR0207379A (pt) | 2004-06-15 |
| CA2439042C (en) | 2013-06-18 |
| EP1364032A1 (en) | 2003-11-26 |
| IL157416A0 (en) | 2004-03-28 |
| IL157416A (en) | 2009-08-03 |
| EP1364032B1 (en) | 2006-10-11 |
| NZ527642A (en) | 2005-03-24 |
| CN1279171C (zh) | 2006-10-11 |
| ZA200305868B (en) | 2005-02-23 |
| KR100858833B1 (ko) | 2008-09-17 |
| KR20030074841A (ko) | 2003-09-19 |
| MXPA03007116A (es) | 2003-11-18 |
| CA2439042A1 (en) | 2002-09-12 |
| JP4199543B2 (ja) | 2008-12-17 |
| CY1106274T1 (el) | 2011-10-12 |
| DE60215315T2 (de) | 2007-05-24 |
| ATE342367T1 (de) | 2006-11-15 |
| WO2002070722A1 (en) | 2002-09-12 |
| PT1364032E (pt) | 2007-01-31 |
| AU2002250903B2 (en) | 2006-12-07 |
| DK1364032T3 (da) | 2007-02-19 |
| ES2272696T3 (es) | 2007-05-01 |
| DE60215315D1 (de) | 2006-11-23 |
| CN1526021A (zh) | 2004-09-01 |
| NO20033672D0 (no) | 2003-08-19 |
| NO20033672L (no) | 2003-10-17 |
| HK1065068A1 (en) | 2005-02-08 |
| JP2004518446A (ja) | 2004-06-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| NO331376B1 (no) | DNA-molekyl kodende for fusjonsproteiner der den N-terminale delen er et hirudinderivat for fremstilling av rekombinante proteiner via sekresjon fra gjaer, samt multikopivektor, plasmid, vertscelle og fremgangsmater for fermentering og fremstilling | |
| CA2438886C (en) | Fusion protein for the secretion of a protein of interest into the supernatant of the bacterial culture | |
| US5962267A (en) | Proinsulin derivative and process for producing human insulin | |
| EP2406377B1 (en) | A polynucleotide and polypeptide sequence and methods thereof | |
| AU2002250903A1 (en) | Use of fusion proteins whose N-termial parts is a hirudin derivative for the production of recombinant proteins via secretion by yeasts | |
| AU2002231784A1 (en) | Fusion protein for the secretion of a protein of interest into the supernatant of the bacterial culture | |
| JP4243103B2 (ja) | 過分泌可能なペプチドの製造、および、１またはそれ以上の他の所定のポリペプチドの輸送形態の同時改善のための方法における、過分泌可能なペプチドの使用 | |
| US7202059B2 (en) | Fusion proteins capable of being secreted into a fermentation medium | |
| AU2002247696A1 (en) | Use of supersecretable peptides in processes for their production and for parallel improvement of the exported forms of one or more other polypeptides | |
| US20160168226A1 (en) | Process for production of insulin and insulin analogues | |
| EP0346500A1 (en) | Elastase inhibiting polypeptides and process for their preparation by genetic recombination | |
| US7638618B2 (en) | Nucleic acids encoding a hirudin and pro-insulin as superscretable peptides and for parallel improvement of the exported forms of one or more polypeptides of interest | |
| KR100451432B1 (ko) | Ｔ-20 펩타이드의 생물학적 제조방법 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MM1K | Lapsed by not paying the annual fees |