NO329246B1 - Fremgangsmate for fremstilling av proteiner ved autoproteolytisk spalting ved bruk av ekspresjonsvektor og bakterievertscelle - Google Patents
Fremgangsmate for fremstilling av proteiner ved autoproteolytisk spalting ved bruk av ekspresjonsvektor og bakterievertscelle Download PDFInfo
- Publication number
- NO329246B1 NO329246B1 NO20020507A NO20020507A NO329246B1 NO 329246 B1 NO329246 B1 NO 329246B1 NO 20020507 A NO20020507 A NO 20020507A NO 20020507 A NO20020507 A NO 20020507A NO 329246 B1 NO329246 B1 NO 329246B1
- Authority
- NO
- Norway
- Prior art keywords
- polypeptide
- pro
- expression vector
- host cell
- fusion protein
- Prior art date
Links
- 238000003776 cleavage reaction Methods 0.000 title claims description 48
- 230000007017 scission Effects 0.000 title claims description 47
- 230000000468 autoproteolytic effect Effects 0.000 title claims description 35
- 230000001580 bacterial effect Effects 0.000 title claims description 33
- 239000013604 expression vector Substances 0.000 title claims description 22
- 238000000034 method Methods 0.000 title claims description 22
- 108090000623 proteins and genes Proteins 0.000 title description 58
- 102000004169 proteins and genes Human genes 0.000 title description 42
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 57
- 229920001184 polypeptide Polymers 0.000 claims description 55
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 55
- 108020001507 fusion proteins Proteins 0.000 claims description 50
- 102000037865 fusion proteins Human genes 0.000 claims description 50
- 150000001413 amino acids Chemical group 0.000 claims description 42
- 210000004027 cell Anatomy 0.000 claims description 37
- 241000710778 Pestivirus Species 0.000 claims description 19
- 238000004519 manufacturing process Methods 0.000 claims description 12
- 210000004899 c-terminal region Anatomy 0.000 claims description 10
- 241000588724 Escherichia coli Species 0.000 claims description 9
- 239000013598 vector Substances 0.000 claims description 9
- 150000007523 nucleic acids Chemical class 0.000 claims description 8
- 239000013612 plasmid Substances 0.000 claims description 8
- 108020004707 nucleic acids Proteins 0.000 claims description 7
- 102000039446 nucleic acids Human genes 0.000 claims description 7
- 241000710780 Bovine viral diarrhea virus 1 Species 0.000 claims description 6
- 108020004414 DNA Proteins 0.000 claims description 3
- 102000053602 DNA Human genes 0.000 claims description 2
- LXWYCLOUQZZDBD-LIYNQYRNSA-N csfv Chemical compound C([C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)[C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LXWYCLOUQZZDBD-LIYNQYRNSA-N 0.000 claims 6
- PPBOKXIGFIBOGK-BDTUAEFFSA-N bvdv Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)C(C)C)[C@@H](C)CC)C1=CN=CN1 PPBOKXIGFIBOGK-BDTUAEFFSA-N 0.000 claims 1
- 235000001014 amino acid Nutrition 0.000 description 29
- 235000018102 proteins Nutrition 0.000 description 27
- 241000710777 Classical swine fever virus Species 0.000 description 13
- 101100338243 Caenorhabditis elegans hil-6 gene Proteins 0.000 description 12
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 10
- 101710181812 Methionine aminopeptidase Proteins 0.000 description 8
- 230000006698 induction Effects 0.000 description 8
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 8
- 229930182817 methionine Natural products 0.000 description 8
- 235000015097 nutrients Nutrition 0.000 description 8
- 108091034117 Oligonucleotide Proteins 0.000 description 7
- 238000012545 processing Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 241001625930 Luria Species 0.000 description 6
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 108020004705 Codon Proteins 0.000 description 5
- 238000007792 addition Methods 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 5
- 229960000723 ampicillin Drugs 0.000 description 5
- 238000010367 cloning Methods 0.000 description 5
- 210000000805 cytoplasm Anatomy 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 239000000411 inducer Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 238000013519 translation Methods 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- 102000014150 Interferons Human genes 0.000 description 4
- 108010050904 Interferons Proteins 0.000 description 4
- 108091005804 Peptidases Proteins 0.000 description 4
- 108010076039 Polyproteins Proteins 0.000 description 4
- 239000004365 Protease Substances 0.000 description 4
- 235000018417 cysteine Nutrition 0.000 description 4
- 125000000151 cysteine group Chemical class N[C@@H](CS)C(=O)* 0.000 description 4
- 239000013613 expression plasmid Substances 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 3
- 108010076504 Protein Sorting Signals Proteins 0.000 description 3
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 3
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 3
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 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 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 230000001131 transforming effect Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 241000724653 Borna disease virus Species 0.000 description 2
- 108010059378 Endopeptidases Proteins 0.000 description 2
- 102000005593 Endopeptidases Human genes 0.000 description 2
- 241000588722 Escherichia Species 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 102000002265 Human Growth Hormone Human genes 0.000 description 2
- 108010000521 Human Growth Hormone Proteins 0.000 description 2
- 239000000854 Human Growth Hormone Substances 0.000 description 2
- 102000004889 Interleukin-6 Human genes 0.000 description 2
- 108090001005 Interleukin-6 Proteins 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 101100084022 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) lapA gene Proteins 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 102400000827 Saposin-D Human genes 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- 101710137500 T7 RNA polymerase Proteins 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000001086 cytosolic effect Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 210000001723 extracellular space Anatomy 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 229940079322 interferon Drugs 0.000 description 2
- 229940047124 interferons Drugs 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 238000013411 master cell bank Methods 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 101150009573 phoA gene Proteins 0.000 description 2
- 230000001323 posttranslational effect Effects 0.000 description 2
- 229960000856 protein c Drugs 0.000 description 2
- 238000001742 protein purification Methods 0.000 description 2
- 230000012743 protein tagging Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000002741 site-directed mutagenesis Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- PLVPPLCLBIEYEA-WAYWQWQTSA-N (z)-3-(1h-indol-3-yl)prop-2-enoic acid Chemical compound C1=CC=C2C(\C=C/C(=O)O)=CNC2=C1 PLVPPLCLBIEYEA-WAYWQWQTSA-N 0.000 description 1
- 102000004400 Aminopeptidases Human genes 0.000 description 1
- 108090000915 Aminopeptidases Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241001118702 Border disease virus Species 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- 241000863013 Caulobacter sp. Species 0.000 description 1
- 241000010804 Caulobacter vibrioides Species 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 108010013369 Enteropeptidase Proteins 0.000 description 1
- 102100029727 Enteropeptidase Human genes 0.000 description 1
- 241000709661 Enterovirus Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000003951 Erythropoietin Human genes 0.000 description 1
- 108090000394 Erythropoietin Proteins 0.000 description 1
- 241001522878 Escherichia coli B Species 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 108010074860 Factor Xa Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 101000930822 Giardia intestinalis Dipeptidyl-peptidase 4 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
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 1
- 102100039619 Granulocyte colony-stimulating factor Human genes 0.000 description 1
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 description 1
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 description 1
- 108010002231 IgA-specific serine endopeptidase Proteins 0.000 description 1
- 102100037852 Insulin-like growth factor I Human genes 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 206010056254 Intrauterine infection Diseases 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 101001055320 Myxine glutinosa Insulin-like growth factor Proteins 0.000 description 1
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 description 1
- 101800000021 N-terminal protease Proteins 0.000 description 1
- 101710144111 Non-structural protein 3 Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 101800001494 Protease 2A Proteins 0.000 description 1
- 102100037681 Protein FEV Human genes 0.000 description 1
- 101710198166 Protein FEV Proteins 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 241000282849 Ruminantia Species 0.000 description 1
- 101800001098 Serine protease NS3 Proteins 0.000 description 1
- 101800001838 Serine protease/helicase NS3 Proteins 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009697 arginine Nutrition 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 238000005844 autocatalytic reaction Methods 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000011210 chromatographic step Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000000120 cytopathologic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 238000011143 downstream manufacturing Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 229940066758 endopeptidases Drugs 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 229940105423 erythropoietin Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 101150047862 hil-6 gene Proteins 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 229940116886 human interleukin-6 Drugs 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- PLVPPLCLBIEYEA-UHFFFAOYSA-N indoleacrylic acid Natural products C1=CC=C2C(C=CC(=O)O)=CNC2=C1 PLVPPLCLBIEYEA-UHFFFAOYSA-N 0.000 description 1
- 239000003262 industrial enzyme Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 229940047122 interleukins Drugs 0.000 description 1
- 235000018977 lysine Nutrition 0.000 description 1
- 230000001320 lysogenic effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000010327 methods by industry Methods 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000004897 n-terminal region Anatomy 0.000 description 1
- 102000034288 naturally occurring fusion proteins Human genes 0.000 description 1
- 108091006048 naturally occurring fusion proteins Proteins 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 210000001322 periplasm Anatomy 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 230000029663 wound healing Effects 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
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- 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
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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)
- C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
- C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/50—Fusion polypeptide containing protease site
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
- C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
- C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Enzymes And Modification Thereof (AREA)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AT136899 | 1999-08-09 | ||
PCT/EP2000/007642 WO2001011056A1 (en) | 1999-08-09 | 2000-08-07 | Production of proteins by autoproteolytic cleavage |
Publications (3)
Publication Number | Publication Date |
---|---|
NO20020507D0 NO20020507D0 (no) | 2002-01-31 |
NO20020507L NO20020507L (no) | 2002-04-05 |
NO329246B1 true NO329246B1 (no) | 2010-09-20 |
Family
ID=3512383
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
NO20020507A NO329246B1 (no) | 1999-08-09 | 2002-01-31 | Fremgangsmate for fremstilling av proteiner ved autoproteolytisk spalting ved bruk av ekspresjonsvektor og bakterievertscelle |
Country Status (17)
Country | Link |
---|---|
US (1) | US7378512B2 (zh) |
EP (1) | EP1200603B1 (zh) |
JP (1) | JP5480458B2 (zh) |
KR (1) | KR100735791B1 (zh) |
CN (1) | CN1230542C (zh) |
AU (1) | AU775681B2 (zh) |
CA (1) | CA2380302C (zh) |
CZ (1) | CZ303341B6 (zh) |
HK (1) | HK1047127B (zh) |
HU (1) | HU230117B1 (zh) |
IL (2) | IL147411A0 (zh) |
MX (1) | MXPA02001425A (zh) |
NO (1) | NO329246B1 (zh) |
PL (1) | PL203708B1 (zh) |
SK (1) | SK288290B6 (zh) |
TR (1) | TR200200048T2 (zh) |
WO (1) | WO2001011056A1 (zh) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2823220B1 (fr) | 2001-04-04 | 2003-12-12 | Genodyssee | Nouveaux polynucleotides et polypeptides de l'erythropoietine (epo) |
JP2005510244A (ja) * | 2001-11-26 | 2005-04-21 | ザ・ユニヴァーシティ・オヴ・クイーンズランド | フラビウイルスワクチン送達系 |
GB0508434D0 (en) * | 2005-04-26 | 2005-06-01 | Sandoz Ag | Organic compounds |
AU2011253661B2 (en) * | 2005-04-26 | 2013-06-13 | Boehringer Ingelheim Rcv Gmbh & Co Kg | Production of recombinant proteins by autoproteolytic cleavage of a fusion protein |
AU2006239721B2 (en) | 2005-04-26 | 2011-07-14 | Boehringer Ingelheim Rcv Gmbh & Co Kg | Production of recombinant proteins by autoproteolytic cleavage of a fusion protein |
GB0508435D0 (en) * | 2005-04-26 | 2005-06-01 | Sandoz Ag | Organic compounds |
US20100137563A1 (en) * | 2008-12-03 | 2010-06-03 | Northwestern University | Cysteine Protease Autoprocessing of Fusion Proteins |
EP2684951A1 (en) | 2012-07-13 | 2014-01-15 | Sandoz Ag | Method for producing a recombinant protein of interest |
EP2746391A1 (en) | 2012-12-19 | 2014-06-25 | Sandoz Ag | Method for producing a recombinant protein of interest |
EP2746390A1 (en) * | 2012-12-19 | 2014-06-25 | Sandoz Ag | Method for producing a recombinant protein of interest |
UY35874A (es) | 2013-12-12 | 2015-07-31 | Novartis Ag | Un proceso para la preparación de una composición de proteínas pegiladas |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4743679A (en) * | 1986-02-24 | 1988-05-10 | Creative Biomolecules, Inc. | Process for producing human epidermal growth factor and analogs thereof |
ES2059482T3 (es) | 1987-12-23 | 1994-11-16 | Boehringer Ingelheim Int | Expresion de la proteasa p2a de hrv2 codificada por virus. |
US6077694A (en) * | 1990-09-21 | 2000-06-20 | The United States Of America As Represented By The Department Of Health And Human Services | Method for over-expression and rapid purification of biosynthetic proteins |
BR9809416A (pt) * | 1997-04-25 | 2000-06-13 | Sembiosys Genetics Inc | Processo para clivagem de proteìnas de fusão |
WO1999010483A2 (de) * | 1997-08-22 | 1999-03-04 | Roche Diagnostics Gmbh | Autokatalytisch aktivierbare vorstufen von proteasen und deren verwendung |
WO1999010503A1 (de) * | 1997-08-22 | 1999-03-04 | Roche Diagnostics Gmbh | Autokatalytisch aktivierbare zymogene vorstufen von proteasen und deren verwendung |
ATE335826T1 (de) * | 1999-08-09 | 2006-09-15 | Sandoz Ag | Proteinherstellung |
-
2000
- 2000-08-07 KR KR1020027001667A patent/KR100735791B1/ko active IP Right Grant
- 2000-08-07 TR TR2002/00048T patent/TR200200048T2/xx unknown
- 2000-08-07 JP JP2001515841A patent/JP5480458B2/ja not_active Expired - Lifetime
- 2000-08-07 SK SK187-2002A patent/SK288290B6/sk not_active IP Right Cessation
- 2000-08-07 WO PCT/EP2000/007642 patent/WO2001011056A1/en active IP Right Grant
- 2000-08-07 CA CA2380302A patent/CA2380302C/en not_active Expired - Lifetime
- 2000-08-07 HU HU0203051A patent/HU230117B1/hu unknown
- 2000-08-07 AU AU66998/00A patent/AU775681B2/en not_active Expired
- 2000-08-07 CZ CZ20020480A patent/CZ303341B6/cs not_active IP Right Cessation
- 2000-08-07 CN CNB008110719A patent/CN1230542C/zh not_active Expired - Lifetime
- 2000-08-07 PL PL353090A patent/PL203708B1/pl unknown
- 2000-08-07 IL IL14741100A patent/IL147411A0/xx unknown
- 2000-08-07 EP EP00954596.3A patent/EP1200603B1/en not_active Expired - Lifetime
- 2000-08-07 MX MXPA02001425A patent/MXPA02001425A/es active IP Right Grant
-
2001
- 2001-12-31 IL IL147411A patent/IL147411A/en active IP Right Grant
-
2002
- 2002-01-31 NO NO20020507A patent/NO329246B1/no not_active IP Right Cessation
- 2002-09-24 HK HK02106953.0A patent/HK1047127B/zh not_active IP Right Cessation
-
2004
- 2004-01-23 US US10/763,619 patent/US7378512B2/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
SK288290B6 (sk) | 2015-07-01 |
JP2003506092A (ja) | 2003-02-18 |
CZ2002480A3 (cs) | 2002-05-15 |
HU230117B1 (hu) | 2015-08-28 |
NO20020507D0 (no) | 2002-01-31 |
HUP0203051A3 (en) | 2003-12-29 |
EP1200603B1 (en) | 2014-12-24 |
US20040215008A1 (en) | 2004-10-28 |
PL353090A1 (en) | 2003-10-06 |
NO20020507L (no) | 2002-04-05 |
MXPA02001425A (es) | 2002-08-12 |
CZ303341B6 (cs) | 2012-08-08 |
US7378512B2 (en) | 2008-05-27 |
PL203708B1 (pl) | 2009-11-30 |
IL147411A0 (en) | 2002-08-14 |
CN1230542C (zh) | 2005-12-07 |
AU775681B2 (en) | 2004-08-12 |
CN1367837A (zh) | 2002-09-04 |
SK1872002A3 (en) | 2002-07-02 |
EP1200603A1 (en) | 2002-05-02 |
IL147411A (en) | 2013-12-31 |
HUP0203051A2 (hu) | 2002-12-28 |
HK1047127A1 (zh) | 2003-02-07 |
TR200200048T2 (tr) | 2002-04-22 |
WO2001011056A1 (en) | 2001-02-15 |
CA2380302C (en) | 2013-02-26 |
HK1047127B (zh) | 2015-08-21 |
KR20020021175A (ko) | 2002-03-18 |
CA2380302A1 (en) | 2001-02-15 |
JP5480458B2 (ja) | 2014-04-23 |
AU6699800A (en) | 2001-03-05 |
KR100735791B1 (ko) | 2007-07-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Dümmler et al. | Simplified screening for the detection of soluble fusion constructs expressed in E. coli using a modular set of vectors | |
US20050186564A1 (en) | Production of proteins | |
KR101374149B1 (ko) | 융합 단백질의 자가단백질분해성 절단에 의한 재조합단백질 생산 | |
EP0682709A1 (en) | Fusion proteins including a cleavage site recognized by a plant virus protease | |
US11053505B2 (en) | Cleavable fusion tag for protein overexpression and purification | |
NO329246B1 (no) | Fremgangsmate for fremstilling av proteiner ved autoproteolytisk spalting ved bruk av ekspresjonsvektor og bakterievertscelle | |
BR9810650B1 (pt) | vetor para expressço de proteÍna heteràloga e mÉtodos para extrair proteÍna recombinante e para purificar insulina recombinante isolada. | |
Bell | E. coli expression systems | |
Whitmarsh et al. | Protein fusions as an aid to purification | |
Han | Control of Intein-Mediated Self-Cleaving Tag for Recombinant Protein Purification | |
Makkapati | Recombinant production of peptides using SUMO as a fusion partner |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
MK1K | Patent expired |