CN100395263C - 缺氧条件下的蛋白质分离方法 - Google Patents
缺氧条件下的蛋白质分离方法 Download PDFInfo
- Publication number
- CN100395263C CN100395263C CNB2004800030976A CN200480003097A CN100395263C CN 100395263 C CN100395263 C CN 100395263C CN B2004800030976 A CNB2004800030976 A CN B2004800030976A CN 200480003097 A CN200480003097 A CN 200480003097A CN 100395263 C CN100395263 C CN 100395263C
- Authority
- CN
- China
- Prior art keywords
- protein
- cell
- inclusion body
- disulfide linkage
- basic
- 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 - Lifetime
Links
- 238000000034 method Methods 0.000 title claims abstract description 72
- 238000000164 protein isolation Methods 0.000 title abstract description 4
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 89
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 89
- 239000002243 precursor Substances 0.000 claims abstract description 7
- 210000004027 cell Anatomy 0.000 claims description 35
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 claims description 27
- 210000003000 inclusion body Anatomy 0.000 claims description 25
- 108090001061 Insulin Proteins 0.000 claims description 21
- 102000004877 Insulin Human genes 0.000 claims description 20
- 230000008569 process Effects 0.000 claims description 14
- 239000012298 atmosphere Substances 0.000 claims description 12
- 238000000926 separation method Methods 0.000 claims description 12
- 230000008521 reorganization Effects 0.000 claims description 4
- 239000012299 nitrogen atmosphere Substances 0.000 claims description 3
- 239000013613 expression plasmid Substances 0.000 claims description 2
- 108010076181 Proinsulin Proteins 0.000 abstract description 11
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 abstract description 10
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 abstract description 6
- 238000002955 isolation Methods 0.000 abstract 2
- 239000003638 chemical reducing agent Substances 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 72
- 108090000765 processed proteins & peptides Proteins 0.000 description 24
- 229920001184 polypeptide Polymers 0.000 description 18
- 102000004196 processed proteins & peptides Human genes 0.000 description 18
- 230000002829 reductive effect Effects 0.000 description 16
- 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 description 15
- 206010002660 Anoxia Diseases 0.000 description 14
- 241000976983 Anoxia Species 0.000 description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- 206010021143 Hypoxia Diseases 0.000 description 14
- 230000007953 anoxia Effects 0.000 description 14
- 239000003795 chemical substances by application Substances 0.000 description 14
- 230000003647 oxidation Effects 0.000 description 9
- 238000007254 oxidation reaction Methods 0.000 description 9
- 229910052760 oxygen Inorganic materials 0.000 description 9
- 229940125396 insulin Drugs 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- CWHJIJJSDGEHNS-MYLFLSLOSA-N Senegenin Chemical compound C1[C@H](O)[C@H](O)[C@@](C)(C(O)=O)[C@@H]2CC[C@@]3(C)C(CC[C@]4(CCC(C[C@H]44)(C)C)C(O)=O)=C4[C@@H](CCl)C[C@@H]3[C@]21C CWHJIJJSDGEHNS-MYLFLSLOSA-N 0.000 description 6
- 238000012545 processing Methods 0.000 description 6
- 239000009871 tenuigenin Substances 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- 102000013090 Thioredoxin-Disulfide Reductase Human genes 0.000 description 5
- 108010079911 Thioredoxin-disulfide reductase Proteins 0.000 description 5
- 238000005336 cracking Methods 0.000 description 5
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 108010075254 C-Peptide Proteins 0.000 description 4
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 4
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 4
- XVOYSCVBGLVSOL-UHFFFAOYSA-N cysteic acid Chemical compound OC(=O)C(N)CS(O)(=O)=O XVOYSCVBGLVSOL-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 3
- 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 3
- 241000282414 Homo sapiens Species 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000009089 cytolysis Effects 0.000 description 3
- 210000000172 cytosol Anatomy 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- PBGKTOXHQIOBKM-FHFVDXKLSA-N insulin (human) Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)[C@@H](C)CC)[C@@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 PBGKTOXHQIOBKM-FHFVDXKLSA-N 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000001742 protein purification Methods 0.000 description 3
- 238000004007 reversed phase HPLC Methods 0.000 description 3
- DEQANNDTNATYII-OULOTJBUSA-N (4r,7s,10s,13r,16s,19r)-10-(4-aminobutyl)-19-[[(2r)-2-amino-3-phenylpropanoyl]amino]-16-benzyl-n-[(2r,3r)-1,3-dihydroxybutan-2-yl]-7-[(1r)-1-hydroxyethyl]-13-(1h-indol-3-ylmethyl)-6,9,12,15,18-pentaoxo-1,2-dithia-5,8,11,14,17-pentazacycloicosane-4-carboxa Chemical compound C([C@@H](N)C(=O)N[C@H]1CSSC[C@H](NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](CC=2C3=CC=CC=C3NC=2)NC(=O)[C@H](CC=2C=CC=CC=2)NC1=O)C(=O)N[C@H](CO)[C@H](O)C)C1=CC=CC=C1 DEQANNDTNATYII-OULOTJBUSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 108050001049 Extracellular proteins Proteins 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 101000976075 Homo sapiens Insulin Proteins 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 108010016076 Octreotide Proteins 0.000 description 2
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000000813 peptide hormone Substances 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000010361 transduction Methods 0.000 description 2
- 230000026683 transduction Effects 0.000 description 2
- DSSYKIVIOFKYAU-XCBNKYQSSA-N (R)-camphor Chemical compound C1C[C@@]2(C)C(=O)C[C@@H]1C2(C)C DSSYKIVIOFKYAU-XCBNKYQSSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 102000003670 Carboxypeptidase B Human genes 0.000 description 1
- 108090000087 Carboxypeptidase B Proteins 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 108010025905 Cystine-Knot Miniproteins Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 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 1
- 241000206602 Eukaryota Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101000911390 Homo sapiens Coagulation factor VIII Proteins 0.000 description 1
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 description 1
- 102000000853 LDL receptors Human genes 0.000 description 1
- 108010001831 LDL receptors Proteins 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- 108010006519 Molecular Chaperones Proteins 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- SCKXCAADGDQQCS-UHFFFAOYSA-N Performic acid Chemical compound OOC=O SCKXCAADGDQQCS-UHFFFAOYSA-N 0.000 description 1
- 102000029797 Prion Human genes 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 102000005157 Somatostatin Human genes 0.000 description 1
- 108010056088 Somatostatin Proteins 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical class N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000003570 biosynthesizing effect Effects 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 229960000846 camphor Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 108050003126 conotoxin Proteins 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- ZUBDGKVDJUIMQQ-ZTNLKOGPSA-N endothelin i Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)NC(=O)[C@H]1NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC=2C=CC(O)=CC=2)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]2CSSC[C@@H](C(N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N2)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CSSC1)C1=CNC=N1 ZUBDGKVDJUIMQQ-ZTNLKOGPSA-N 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 102000057593 human F8 Human genes 0.000 description 1
- 201000001421 hyperglycemia Diseases 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229950004152 insulin human Drugs 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 229960003151 mercaptamine Drugs 0.000 description 1
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229960002700 octreotide Drugs 0.000 description 1
- 229910000073 phosphorus hydride Inorganic materials 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 229940047431 recombinate Drugs 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229940072272 sandostatin Drugs 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000002795 scorpion venom Substances 0.000 description 1
- 238000009991 scouring Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 1
- 229960000553 somatostatin Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 210000004895 subcellular structure Anatomy 0.000 description 1
- 238000006277 sulfonation reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- CWERGRDVMFNCDR-UHFFFAOYSA-N thioglycolic acid Chemical compound OC(=O)CS CWERGRDVMFNCDR-UHFFFAOYSA-N 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- -1 vassopressin Proteins 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
- C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
Description
Claims (10)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP03100208.2 | 2003-01-31 | ||
EP03100208 | 2003-01-31 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1745092A CN1745092A (zh) | 2006-03-08 |
CN100395263C true CN100395263C (zh) | 2008-06-18 |
Family
ID=32799004
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB2004800030976A Expired - Lifetime CN100395263C (zh) | 2003-01-31 | 2004-01-29 | 缺氧条件下的蛋白质分离方法 |
Country Status (11)
Country | Link |
---|---|
US (3) | US20060128942A1 (zh) |
EP (1) | EP1592703B1 (zh) |
JP (1) | JP4992029B2 (zh) |
CN (1) | CN100395263C (zh) |
AT (1) | ATE397619T1 (zh) |
CA (1) | CA2514416C (zh) |
DE (1) | DE602004014251D1 (zh) |
ES (1) | ES2306981T3 (zh) |
HK (1) | HK1078885A1 (zh) |
PL (1) | PL212317B1 (zh) |
WO (1) | WO2004067556A1 (zh) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004067556A1 (en) * | 2003-01-31 | 2004-08-12 | Akzo Nobel N.V. | Method for protein isolation in anoxic conditions |
US20120214199A1 (en) * | 2011-02-23 | 2012-08-23 | Elona Biotechnologies | Lis-pro proinsulin compositions and methods of producing lis-pro insulin analogs therefrom |
MX354993B (es) | 2007-07-09 | 2018-03-28 | Genentech Inc | Prevención de reducción de enlaces de disulfuro durante la producción recombinante de polipéptidos. |
CN101782549A (zh) * | 2010-03-29 | 2010-07-21 | 内蒙古蒙牛乳业(集团)股份有限公司 | 检测牛乳制品中酪蛋白磷酸肽含量的方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1290299A (zh) * | 1997-12-29 | 2001-04-04 | 株式会社钟根堂 | 人胰岛素原的制备方法 |
Family Cites Families (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
NZ199391A (en) | 1981-01-02 | 1985-12-13 | Genentech Inc | Chimeric polypeptides comprising a proinsulin sequence,and preparation by recombinant dna technique;production of human insulin |
CA1260858A (en) | 1985-03-28 | 1989-09-26 | Lawrence S. Cousens | Expression using fused genes providing for protein product |
US4801691A (en) * | 1987-05-15 | 1989-01-31 | International Minerals & Chemical Corp. | Method for removing sodium dodecyl sulfate from sodium dodecyl sulfate solubilized protein solutions |
US5593860A (en) * | 1987-08-14 | 1997-01-14 | Bio-Technology General Corp. | Expression plasmids containing the deo promoter, and bacterial hosts containing the plasmids |
EP0347781B1 (de) * | 1988-06-23 | 1994-02-16 | Hoechst Aktiengesellschaft | Mini-Proinsulin, seine Herstellung und Verwendung |
US6875589B1 (en) * | 1988-06-23 | 2005-04-05 | Hoechst Aktiengesellschaft | Mini-proinsulin, its preparation and use |
US4923967A (en) * | 1988-09-26 | 1990-05-08 | Eli Lilly And Company | Purification and refolding of recombinant proteins |
DE3901718A1 (de) | 1989-01-21 | 1990-07-26 | Hoechst Ag | Verfahren zur renaturierung falscher rekombinanten von insulinvorlaeufern |
JP2857684B2 (ja) * | 1989-05-04 | 1999-02-17 | ジェネンテク,インコーポレイテッド | ヒトリラキシンの単離のための方法および組成物 |
US6020145A (en) * | 1989-06-30 | 2000-02-01 | Bristol-Myers Squibb Company | Methods for determining the presence of carcinoma using the antigen binding region of monoclonal antibody BR96 |
US5840851A (en) * | 1993-07-23 | 1998-11-24 | Plomer; J. Jeffrey | Purification of hemoglobin |
US6001604A (en) * | 1993-12-29 | 1999-12-14 | Bio-Technology General Corp. | Refolding of proinsulins without addition of reducing agents |
JPH09503396A (ja) * | 1994-07-29 | 1997-04-08 | インノジェネティクス・エヌ・ブイ | 診断用及び治療用の精製c型肝炎ウイルスエンベロープ蛋白 |
HU223718B1 (hu) | 1994-12-29 | 2004-12-28 | Savient Pharmaceuticals, Inc. | Eljárás humán inzulin előállítására, proinzulin-tartalmú hibridpolipeptid expressziójából kiindulva |
WO2004067556A1 (en) * | 2003-01-31 | 2004-08-12 | Akzo Nobel N.V. | Method for protein isolation in anoxic conditions |
UA96139C2 (uk) * | 2005-11-08 | 2011-10-10 | Дженентек, Інк. | Антитіло до нейропіліну-1 (nrp1) |
-
2004
- 2004-01-29 WO PCT/EP2004/000953 patent/WO2004067556A1/en active IP Right Grant
- 2004-01-29 PL PL377927A patent/PL212317B1/pl unknown
- 2004-01-29 CN CNB2004800030976A patent/CN100395263C/zh not_active Expired - Lifetime
- 2004-01-29 DE DE602004014251T patent/DE602004014251D1/de not_active Expired - Lifetime
- 2004-01-29 EP EP04706187A patent/EP1592703B1/en not_active Expired - Lifetime
- 2004-01-29 AT AT04706187T patent/ATE397619T1/de not_active IP Right Cessation
- 2004-01-29 JP JP2006501710A patent/JP4992029B2/ja not_active Expired - Lifetime
- 2004-01-29 US US10/543,524 patent/US20060128942A1/en not_active Abandoned
- 2004-01-29 ES ES04706187T patent/ES2306981T3/es not_active Expired - Lifetime
- 2004-01-29 CA CA2514416A patent/CA2514416C/en not_active Expired - Lifetime
-
2006
- 2006-02-07 HK HK06101615A patent/HK1078885A1/xx not_active IP Right Cessation
-
2012
- 2012-05-09 US US13/467,305 patent/US8802395B2/en not_active Expired - Lifetime
-
2014
- 2014-07-08 US US14/325,822 patent/US9505802B2/en not_active Expired - Lifetime
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1290299A (zh) * | 1997-12-29 | 2001-04-04 | 株式会社钟根堂 | 人胰岛素原的制备方法 |
Non-Patent Citations (2)
Title |
---|
Protein folding. R. Hermann.European Patent Office. 1993 |
Protein folding. R. Hermann.European Patent Office. 1993 * |
Also Published As
Publication number | Publication date |
---|---|
WO2004067556A1 (en) | 2004-08-12 |
US20130150553A1 (en) | 2013-06-13 |
HK1078885A1 (en) | 2006-03-24 |
PL377927A1 (pl) | 2006-02-20 |
JP4992029B2 (ja) | 2012-08-08 |
US9505802B2 (en) | 2016-11-29 |
US8802395B2 (en) | 2014-08-12 |
EP1592703B1 (en) | 2008-06-04 |
CA2514416A1 (en) | 2004-08-12 |
US20140323687A1 (en) | 2014-10-30 |
ATE397619T1 (de) | 2008-06-15 |
CA2514416C (en) | 2012-08-14 |
PL212317B1 (pl) | 2012-09-28 |
JP2007506406A (ja) | 2007-03-22 |
EP1592703A1 (en) | 2005-11-09 |
DE602004014251D1 (de) | 2008-07-17 |
CN1745092A (zh) | 2006-03-08 |
US20060128942A1 (en) | 2006-06-15 |
ES2306981T3 (es) | 2008-11-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Clark | Protein refolding for industrial processes | |
Singh et al. | Solubilization and refolding of bacterial inclusion body proteins | |
DE69434919T2 (de) | Faltung von Polypeptiden | |
US6399333B1 (en) | Process for producing erythropoietin containing no animal proteins | |
AU7561694A (en) | Aqueous multiple-phase isolation of polypeptide | |
IL268868A (en) | A new method of protein cleansing | |
US20030166062A1 (en) | Methods and compositions for production of recombinant peptides | |
US9505802B2 (en) | Method for protein isolation in anoxic conditions | |
EP0248618A2 (en) | Isolation of bioactive, monomeric growth hormone | |
US6916914B1 (en) | Purification of somatotropin from transformed microorganisms | |
US6339061B1 (en) | Process for the stabilization of proteins in complex mixtures during their storage in aqueous solvents | |
McGregor et al. | Large Scale Production of Human Alpha Interferon from Bacteria | |
NO864206L (no) | Derivat av interleukin-2, dets fremstilling og anvendelse. | |
Chaudhuri | NEW DEVELOPMENTS IN THE REFOLDING OF RECOMBINANT PROTEINS | |
MXPA05012298A (es) | Proceso de manufactura de una proteina anticoagulante extractada de nematodos (nap). | |
Phue et al. | Improved refolding of recombinant human proinsulin from Escherichia coli in a two-stage reactor system | |
Ward | Recovery of Cell Products | |
JPH02117395A (ja) | 菌体内からのヒトインターロイキン3様ポリペプチドの回収法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
ASS | Succession or assignment of patent right |
Owner name: ORGANON NV Free format text: FORMER OWNER: AKZO NOVEL N.V. CORP. Effective date: 20061229 |
|
C41 | Transfer of patent application or patent right or utility model | ||
TA01 | Transfer of patent application right |
Effective date of registration: 20061229 Address after: Holland Applicant after: N.V. ORGANON Address before: Holland Arnhem Applicant before: AKZO NOBEL N.V. |
|
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C56 | Change in the name or address of the patentee |
Owner name: MSD OS CO., LTD. Free format text: FORMER NAME: ORGANON NV |
|
CP01 | Change in the name or title of a patent holder |
Address after: Holland Patentee after: N.V. Organon Address before: Holland Patentee before: N.V. ORGANON |
|
ASS | Succession or assignment of patent right |
Owner name: MERCK SHARP + DOHME CORP. Free format text: FORMER OWNER: ORGANON BIOLOGICAL NETHERLANDS CO., LTD. Effective date: 20131011 Owner name: ORGANON BIOLOGICAL NETHERLANDS CO., LTD. Free format text: FORMER OWNER: MSD OUSI CO., LTD. Effective date: 20131011 |
|
C41 | Transfer of patent application or patent right or utility model | ||
TR01 | Transfer of patent right |
Effective date of registration: 20131011 Address after: Haarlem Patentee after: MERCK SHARP & DOHME B.V. Address before: Haarlem Patentee before: Organon biological science Holland Co.,Ltd. Effective date of registration: 20131011 Address after: Haarlem Patentee after: Organon biological science Holland Co.,Ltd. Address before: Holland Patentee before: N.V. Organon |
|
CX01 | Expiry of patent term | ||
CX01 | Expiry of patent term |
Granted publication date: 20080618 |