JP5782580B2 - ポリペプチドヒドロゲル及びその製造方法 - Google Patents
ポリペプチドヒドロゲル及びその製造方法 Download PDFInfo
- Publication number
- JP5782580B2 JP5782580B2 JP2015513725A JP2015513725A JP5782580B2 JP 5782580 B2 JP5782580 B2 JP 5782580B2 JP 2015513725 A JP2015513725 A JP 2015513725A JP 2015513725 A JP2015513725 A JP 2015513725A JP 5782580 B2 JP5782580 B2 JP 5782580B2
- Authority
- JP
- Japan
- Prior art keywords
- polypeptide
- solution
- hydrogel
- water
- solvent
- 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.)
- Active
Links
- 229920001184 polypeptide Polymers 0.000 title claims description 74
- 108090000765 processed proteins & peptides Proteins 0.000 title claims description 74
- 102000004196 processed proteins & peptides Human genes 0.000 title claims description 74
- 239000000017 hydrogel Substances 0.000 title claims description 41
- 238000004519 manufacturing process Methods 0.000 title claims description 14
- 108090000623 proteins and genes Proteins 0.000 claims description 62
- 102000004169 proteins and genes Human genes 0.000 claims description 49
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 46
- 239000002904 solvent Substances 0.000 claims description 32
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 27
- 229920001872 Spider silk Polymers 0.000 claims description 26
- 239000003021 water soluble solvent Substances 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 238000004090 dissolution Methods 0.000 claims description 11
- 238000000502 dialysis Methods 0.000 claims description 8
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 6
- 238000006467 substitution reaction Methods 0.000 claims description 6
- 150000001413 amino acids Chemical group 0.000 description 44
- 235000018102 proteins Nutrition 0.000 description 43
- 239000000243 solution Substances 0.000 description 42
- 239000000499 gel Substances 0.000 description 17
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 101150082901 ADF3 gene Proteins 0.000 description 7
- 241000239290 Araneae Species 0.000 description 7
- 206010036790 Productive cough Diseases 0.000 description 6
- 108091081024 Start codon Proteins 0.000 description 6
- 239000013078 crystal Substances 0.000 description 6
- 210000003802 sputum Anatomy 0.000 description 6
- 208000024794 sputum Diseases 0.000 description 6
- 108010091324 3C proteases Proteins 0.000 description 5
- 241000430519 Human rhinovirus sp. Species 0.000 description 5
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 108091005804 Peptidases Proteins 0.000 description 5
- 239000004365 Protease Substances 0.000 description 5
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 5
- 235000004279 alanine Nutrition 0.000 description 5
- 238000009835 boiling Methods 0.000 description 5
- 210000000845 cartilage Anatomy 0.000 description 5
- 239000013604 expression vector Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 230000003252 repetitive effect Effects 0.000 description 5
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
- 108010022355 Fibroins Proteins 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 101710112083 Para-Rep C1 Proteins 0.000 description 4
- 102100022881 Rab proteins geranylgeranyltransferase component A 1 Human genes 0.000 description 4
- 101710119887 Trans-acting factor B Proteins 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 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 3
- 229960000723 ampicillin Drugs 0.000 description 3
- 238000012377 drug delivery Methods 0.000 description 3
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 3
- 239000013600 plasmid vector Substances 0.000 description 3
- 238000003752 polymerase chain reaction Methods 0.000 description 3
- 239000012460 protein solution Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000003357 wound healing promoting agent Substances 0.000 description 3
- QSLUMWFISVQCLL-UHFFFAOYSA-N 1,2-dichloroethane;formic acid Chemical compound OC=O.ClCCCl QSLUMWFISVQCLL-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 229920002972 Acrylic fiber Polymers 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 101710084218 Master replication protein Proteins 0.000 description 2
- 101000708578 Milk vetch dwarf virus (isolate N) Para-Rep C3 Proteins 0.000 description 2
- 241000238902 Nephila clavipes Species 0.000 description 2
- 101710112078 Para-Rep C2 Proteins 0.000 description 2
- 102100022880 Rab proteins geranylgeranyltransferase component A 2 Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 101710119961 Trans-acting factor C Proteins 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000010419 fine particle Substances 0.000 description 2
- 210000003495 flagella Anatomy 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- VBZWSGALLODQNC-UHFFFAOYSA-N hexafluoroacetone Chemical compound FC(F)(F)C(=O)C(F)(F)F VBZWSGALLODQNC-UHFFFAOYSA-N 0.000 description 2
- AMXOYNBUYSYVKV-UHFFFAOYSA-M lithium bromide Chemical compound [Li+].[Br-] AMXOYNBUYSYVKV-UHFFFAOYSA-M 0.000 description 2
- 239000008176 lyophilized powder Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000002798 polar solvent Substances 0.000 description 2
- 229920001721 polyimide Polymers 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000013557 residual solvent Substances 0.000 description 2
- VGTPCRGMBIAPIM-UHFFFAOYSA-M sodium thiocyanate Chemical compound [Na+].[S-]C#N VGTPCRGMBIAPIM-UHFFFAOYSA-M 0.000 description 2
- 239000013598 vector Substances 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
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 description 1
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 101150003973 ADF4 gene Proteins 0.000 description 1
- 241000193935 Araneus diadematus Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000196324 Embryophyta Species 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
- VPZXBVLAVMBEQI-VKHMYHEASA-N Glycyl-alanine Chemical compound OC(=O)[C@H](C)NC(=O)CN VPZXBVLAVMBEQI-VKHMYHEASA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 1
- 241000238903 Nephila Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 208000025174 PANDAS Diseases 0.000 description 1
- 208000021155 Paediatric autoimmune neuropsychiatric disorders associated with streptococcal infection Diseases 0.000 description 1
- 240000004718 Panda Species 0.000 description 1
- 235000016496 Panda oleosa Nutrition 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004642 Polyimide Substances 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 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
- 229910001508 alkali metal halide Inorganic materials 0.000 description 1
- 150000008045 alkali metal halides Chemical class 0.000 description 1
- 229910001615 alkaline earth metal halide Inorganic materials 0.000 description 1
- 229910001964 alkaline earth metal nitrate Inorganic materials 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 238000005312 nonlinear dynamic Methods 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 108010054442 polyalanine Proteins 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000009719 polyimide resin Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000012779 reinforcing material Substances 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000005070 sphincter Anatomy 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- 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
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
- C07K14/43518—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from spiders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/008—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/145—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Dermatology (AREA)
- Transplantation (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Botany (AREA)
- Organic Chemistry (AREA)
- Insects & Arthropods (AREA)
- Pharmacology & Pharmacy (AREA)
- Gastroenterology & Hepatology (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- General Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Surgery (AREA)
- Vascular Medicine (AREA)
- Heart & Thoracic Surgery (AREA)
- Materials Engineering (AREA)
- Peptides Or Proteins (AREA)
- Materials For Medical Uses (AREA)
Description
(A)ジメチルスルホキシド
(B)ジメチルスルホキシドに無機塩を加えたもの
(C)N,N−ジメチルホルムアミドに無機塩を加えたもの
前記溶液生成工程で生成した溶液を水溶性溶媒に置換する置換工程を含むことを特徴とする。なお、ここで言う「ポリペプチドを溶解用溶媒に溶解させる」とは、ポリペプチドを溶解用溶媒に完全に溶解させることと、ポリペプチドの微粒子を溶解用溶媒中に分散させることで、ポリペプチドの微粒子を溶解用溶媒に実質的に溶解させた形態と為すことの両方を含む。以下、同一の意味において使用する。
(A)ジメチルスルホキシド
(B)ジメチルスルホキシドに無機塩を加えたもの
(C)N,N−ジメチルホルムアミドに無機塩を加えたもの
前記溶液生成工程で生成した溶液を水溶性溶媒に置換する工程により作成できる。前記溶液生成工程と前記溶解用溶媒を水溶性溶媒に置換する工程の間に、型枠に流し込み所定の形状に成形する工程を入れるか、あるいは前記溶解用溶媒を水溶性溶媒に置換する工程の後にカットすることにより所定の形状とすることができる。得られたポリペプチドヒドロゲルの内部にはジメチルスルホキシド及びN,N−ジメチルホルムアミドからなる群から選ばれる少なくとも一つが存在していてもよい。前記溶解用溶媒の存在量は、特に限定されるものではなく、前記溶液生成工程で生成される溶液を前記水溶性溶媒に置換した後において意図せずに残存する程度の量である。
(1)溶媒残量測定
内部標準として1,2−ジクロロエタン−ギ酸溶液 濃度3,100ppm(0.00310mg/ml)を準備した。タンパク質溶液(10mlのギ酸に0.1gのポリペプチドヒドロゲルを溶解したもの)500μlと内部標準溶液500μlを混合した。さらに、H−NMR測定のためアセトニトリル重溶媒を同量程度加え約2倍に希釈し、H−NMR測定を行った(NMRの機種:JEOL社 JNM−ECX 100)。内部標準試料1,2−ジクロロエタンのH−NMR積分強度とDMSOのH−NMR積分強度を比較した。検量線の作成は3ppm〜3000ppmのDMSO−ギ酸溶液を作成し、上記プロトコルにしたがって、検量線を作成した。検量線との比較から、タンパク質溶液中のDMSO濃度を求めた。DMSO濃度測定は、JEOL社製、核磁気共鳴装置(NMR)を用いた。
(2)粘度
KEM社製、EMS装置を使用した。
(3)水分率
本発明手法に従って製造されたヒドロゲルの質量:W1と、かかるヒドロゲルを完全乾燥させた乾燥体の質量:W2とを測定し、それらの測定値を下記式1に代入して求めた。
水分率=[(W1−W2)×100]/W1・・・式1
1.ポリペプチドの準備
<ADF3Kai−Aの遺伝子の合成>
ニワオニグモの2つの主要なしおり糸タンパク質の一つであるADF3の部分的なアミノ酸配列をNCBIのウェブデータベース(NCBIアクセッション番号:AAC47010、GI:1263287)より取得し、同配列のN末端に開始コドンと、His10タグ及びHRV3Cプロテアーゼ(Human rhinovirus 3Cプロテアーゼ)認識サイトからなるアミノ酸配列(配列番号5)を付加し、該配列のN末端の1残基目から631残基目までのアミノ酸配列(配列番号1)からなるポリペプチド(ADF3Kai−A)をコードする遺伝子を合成した。その結果、配列番号6に示す塩基配列からなるADF3Kai−Aの遺伝子が導入されたpUC57ベクター(遺伝子の5’末端直上流にNde Iサイト、及び5’末端直下流にXba Iサイト有り)を取得した。その後、同遺伝子をNde I及びEcoR Iで制限酵素処理し、pET22b(+)発現ベクターに組み換え、ADF3Kai−Aの遺伝子が導入されたpET22b(+)ベクターを得た。
前記で得られたADF3Kai−Aの遺伝子配列を含むpET22b(+)発現ベクターを、大腸菌Rosetta(DE3)に形質転換した。得られたシングルコロニーを、アンピシリンを含む2mLのLB培地で15時間培養後、同培養液1.4mlを、アンピシリンを含む140mLのLB培地に添加し、37℃、200rpmの条件下で、培養液のOD600が3.5になるまで培養した。次に、OD600が3.5の培養液を、アンピシリンを含む7Lの2×YT培地に50%グルコース140mLと共に加え、OD600が4.0になるまでさらに培養した。その後、得られたOD600が4.0の培養液に、終濃度が0.5mMになるようにイソプロピル−β−チオガラクトピラノシド(IPTG)を添加してタンパク質発現を誘導した。IPTG添加後2時間経過した時点で、培養液を遠心分離し、菌体を回収した。IPTG添加前とIPTG添加後の培養液から調製したタンパク質溶液をポリアクリルアミドゲルに泳動させたところ、IPTG添加に依存して目的サイズ(約56.1kDa)のバンドが観察され、目的とするタンパク質(ADF3Kai−A)が発現していることを確認した。
(1)遠沈管(1000ml)にADF3Kai−Aのタンパク質を発現している大腸菌の菌体約50gと、緩衝液AI(20mM Tris−HCl、pH7.4)300mlを添加し、ミキサー(IKA社製「T18ベーシック ウルトラタラックス」、レベル2)で菌体を分散させた後、遠心分離機(クボタ社製の「Model 7000」)で遠心分離(11,000g、10分、室温)し、上清を捨てた。
(2)遠心分離で得られた沈殿物(菌体)に緩衝液AIを300mlと、0.1MのPMSF(イソプロパノールで溶解)を3ml添加し、前記IKA社製のミキサー(レベル2)で3分間分散させた。その後、高圧ホモジナイザー(GEA Niro Saovi社製の「Panda Plus 2000」)を用いて菌体を繰り返し3回破砕した。
(3)破砕された菌体に、3w/v%のSDSを含む緩衝液B(50mM TrisーHCL、100mM NaCl、pH7.0)300mLを加え、前記IKA社製のミキサー(レベル2)で良く分散させた後、シェイカー(タイテック社製、200rpm、37℃)で60分間攪拌した。その後、前記クボタ社製の遠心分離機で遠心分離(11,000g、30分、室温)し、上清を捨て、SDS洗浄顆粒(沈殿物)を得た。
(4)SDS洗浄顆粒を100mg/mLの濃度になるよう1Mの塩化リチウムを含むDMSO溶液で懸濁し、80℃で1時間熱処理した。その後、前記クボタ社製の遠心分離機で遠心分離(11,000g、30分、室温)し、上清を回収した。
(5)回収した上清に対して3倍量のエタノールを準備し、エタノールに回収した上清を加え、室温で1時間静置した。その後、前記クボタ社製の遠心分離機で遠心分離(11,000g、30分、室温)し、凝集タンパク質を回収した。次に純水を用いて凝集タンパク質を洗浄し、遠心分離により凝集タンパク質を回収するという工程を3回繰り返した後、凍結乾燥機で水分を除き、凍結乾燥粉末を回収した。得られた凍結乾燥粉末における目的タンパク質ADF3Kai−A(約56.1kDa)の精製度は、粉末のポリアクリルアミドゲル電気泳動(CBB染色)の結果をTotallab(nonlinear dynamics ltd.)を用いて画像解析することにより確認した。その結果、ADF3Kai−Aの精製度は約85%であった。
クモ糸タンパク質(ADF3Kai−A)の粉末0.8gをDMSO(1MのLiClを含む)20mlに入れ、80℃で30分間溶解させた。その後、ゴミと泡を取り除いた。溶液の溶液粘度は30.8cP(センチポアズ)であった。この溶液を透析チューブ(三光純薬株式会社セルロースチューブ36/32)に入れた。
前記透析チューブを3リットルの純水で満たされたビーカーに入れた。その後3時間静置させた後、水を入れ替え、計6回この操作を行った。これにより、溶液中のクモ糸タンパク質が凝集して、ほぼ全てのDMSOが水に置換されたヒドロゲルが作成できた。
(1)得られたヒドロゲルの溶媒残量はヒドロゲル100gに対して、0.094gであった。
(2)得られたヒドロゲルの水分率は95.3質量%であった。
(3)得られたヒドロゲルの側面写真を図1に示す。
実施例1と同一のポリペプチドを用い、操作も同様にして次のゲルを作製した。濃度の薄いゲルとして、ポリペプチド粉末0.4gを用いて20mg/mlのドープ(DMSO+1MのLiCl)を準備し、ゲルを作製した。ドープ粘度は13.9cPであった。得られたゲルの水分率は98.8質量%であった。
実施例1と同一のポリペプチドを用い、操作も同様にして次のゲルを作製した。ポリペプチド粉末0.4gを用いて20mg/mlのドープ(塩なしのDMSO)を準備し、ゲルを作製した。得られたゲルの水分率は97.4質量%であった。
実施例1と同一のポリペプチドを用い、操作も同様にして次のゲルを作製した。ポリペプチド粉末0.4gを用いて20mg/mlのドープ(DMF+1MのLiCl)を準備し、ゲルを作製した。得られたゲルの水分率は98.2質量%であった。
配列番号6 塩基配列
Claims (5)
- クモ糸タンパク質に由来するポリペプチドのヒドロゲルであって、前記ポリペプチドヒドロゲル100質量%に対して、水分率が85.0〜99.9質量%であり、
前記ポリペプチドヒドロゲルの内部にはジメチルスルホキシド及びN,N−ジメチルホルムアミドからなる群から選ばれる少なくとも一つが存在していることを特徴とするポリペプチドヒドロゲル。 - クモ糸タンパク質に由来するポリペプチドを、下記(A)〜(C)からなる群から選ばれる少なくとも一つの溶解用溶媒に溶解させてポリペプチドの溶液を得る溶液生成工程と、
(A)ジメチルスルホキシド
(B)ジメチルスルホキシドに無機塩を加えたもの
(C)N,N−ジメチルホルムアミドに無機塩を加えたもの
前記溶液生成工程で生成した溶液を水溶性溶媒に置換する置換工程を含むことを特徴とするポリペプチドヒドロゲルの製造方法。 - 前記置換工程は、前記ポリペプチドの溶液を水に置換する請求項2に記載のポリペプチドヒドロゲルの製造方法。
- 前記置換工程は、前記溶解用溶媒に溶解させたポリペプチドの溶液を透析膜内に入れ、水溶性溶媒中に浸漬し、水溶性溶媒を1回以上入れ替える請求項2又は3に記載のポリペプチドヒドロゲルの製造方法。
- 前記ポリペプチドヒドロゲルは、前記ポリペプチドヒドロゲル100質量%に対して、水分率が85.0〜99.9質量%である請求項2〜4のいずれかに記載のポリペプチドヒドロゲルの製造方法。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2015513725A JP5782580B2 (ja) | 2013-04-25 | 2014-04-18 | ポリペプチドヒドロゲル及びその製造方法 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2013092847 | 2013-04-25 | ||
JP2013092847 | 2013-04-25 | ||
PCT/JP2014/061041 WO2014175177A1 (ja) | 2013-04-25 | 2014-04-18 | ポリペプチドヒドロゲル及びその製造方法 |
JP2015513725A JP5782580B2 (ja) | 2013-04-25 | 2014-04-18 | ポリペプチドヒドロゲル及びその製造方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP5782580B2 true JP5782580B2 (ja) | 2015-09-24 |
JPWO2014175177A1 JPWO2014175177A1 (ja) | 2017-02-23 |
Family
ID=51791755
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015513725A Active JP5782580B2 (ja) | 2013-04-25 | 2014-04-18 | ポリペプチドヒドロゲル及びその製造方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US9968682B2 (ja) |
EP (1) | EP2990062B1 (ja) |
JP (1) | JP5782580B2 (ja) |
CN (1) | CN104936625A (ja) |
WO (1) | WO2014175177A1 (ja) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020067572A1 (ja) | 2018-09-28 | 2020-04-02 | Spiber株式会社 | タンパク質組成物の製造方法 |
WO2021065794A1 (ja) | 2019-09-30 | 2021-04-08 | Spiber株式会社 | タンパク質成形体の製造方法 |
WO2021066078A1 (ja) | 2019-09-30 | 2021-04-08 | Spiber株式会社 | 筋組織再生剤 |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014175178A1 (ja) | 2013-04-25 | 2014-10-30 | スパイバー株式会社 | ポリペプチド多孔質体及びその製造方法 |
CN107735406A (zh) * | 2015-04-09 | 2018-02-23 | 丝芭博株式会社 | 极性溶剂溶液及其制造方法 |
US20180080147A1 (en) | 2015-04-09 | 2018-03-22 | Spiber Inc. | Polar solvent solution and production method thereof |
JP2018159137A (ja) | 2015-08-20 | 2018-10-11 | 国立研究開発法人理化学研究所 | クモ糸様構造を有するポリペプチド繊維の製造方法 |
US20210214404A1 (en) * | 2018-07-25 | 2021-07-15 | Spiber Inc. | Artificial Hair Fiber, Method for Manufacturing Same, and Artificial Hair |
CN110028551A (zh) * | 2019-04-18 | 2019-07-19 | 福州大学 | 一种多肽水凝胶及其制备方法 |
CN117384246B (zh) * | 2023-12-11 | 2024-05-10 | 北京大学第三医院(北京大学第三临床医学院) | 一种多肽及含该多肽的水凝胶、其制备方法与应用 |
Family Cites Families (40)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4448750A (en) | 1981-06-05 | 1984-05-15 | Fuesting Michael L | Sterilization method |
JPH0694518B2 (ja) | 1987-11-02 | 1994-11-24 | 工業技術院長 | 絹フィブロイン多孔質体の製造方法 |
JPH02240165A (ja) | 1989-03-13 | 1990-09-25 | Kanebo Ltd | 保存安定性に優れた絹フィブロイン水溶液及びその製造法 |
JP2940033B2 (ja) | 1989-12-07 | 1999-08-25 | ソニー株式会社 | 走査軌跡評定装置 |
EP0452925B1 (en) | 1990-04-20 | 2000-02-09 | The University Of Wyoming | Isolated DNA coding for spider silk protein, a replicable vector and a transformed cell containing the isolated DNA, and products thereof |
US5989894A (en) | 1990-04-20 | 1999-11-23 | University Of Wyoming | Isolated DNA coding for spider silk protein, a replicable vector and a transformed cell containing the DNA |
US5347754A (en) | 1993-05-10 | 1994-09-20 | Landis & Gyr Powers, Inc. | Position sensing apparatus |
JP2615440B2 (ja) | 1995-01-27 | 1997-05-28 | 農林水産省蚕糸・昆虫農業技術研究所長 | 絹フィブロイン微粉末の製造方法 |
JP4095704B2 (ja) | 1998-02-04 | 2008-06-04 | 出光興産株式会社 | シルク含有樹脂組成物、シルク含有成形品及びシルク含有樹脂組成物の製造方法 |
ATE461996T1 (de) | 1999-09-03 | 2010-04-15 | Univ Ramot | Verbindungen, zusammensetzungen und verfahren zur behandlung oder vorsorge von alzheimer erkrankung |
US6620917B1 (en) | 2000-01-20 | 2003-09-16 | The United States Of America As Represented By The Secretary Of The Army | Method for the purification and aqueous fiber spinning of spider silks and other structural proteins |
BR0111125A (pt) | 2000-05-23 | 2004-12-28 | Cenes Pharmaceuticals Inc | Moléculas de ácido nucléico de nrg-2, polipeptìdeos, e métodos diagnóstico e terapêuticos |
AUPR522501A0 (en) | 2001-05-25 | 2001-06-14 | Proteome Systems Ltd | Increased solubilisation of hydrophobic proteins |
WO2004062697A2 (en) | 2003-01-07 | 2004-07-29 | Tufts University | Silk fibroin materials and use thereof |
EP1613796B1 (en) | 2003-04-10 | 2017-03-22 | Tufts University | Concentrated aqueous silk fibroin solution and use thereof |
EP1660013A4 (en) | 2003-08-26 | 2011-07-20 | Gel Del Technologies Inc | BIOMATERIAL AND PROTEIN BIOCOACERVATES, METHODS OF MAKING AND USING THEM |
CN1243059C (zh) | 2004-03-04 | 2006-02-22 | 苏州大学 | 纳米丝素颗粒的制造方法 |
WO2005124011A1 (en) | 2004-06-10 | 2005-12-29 | Keraplast Technologies, Ltd. | Keratin based hydrogel sheets prepared from fabric for biomedical and other applications and method of production |
RU2415938C2 (ru) | 2004-07-22 | 2011-04-10 | АЭмЗильк ГмбХ | Рекомбинантный белок шелка пауков, кодирующая его нуклеиновая кислота, вектор и клетка-хозяин, подходящие для его экспрессии, способ его агрегации, способ получения материала из него, содержащий его продукт и применение |
RU2421463C2 (ru) | 2005-08-29 | 2011-06-20 | АЭмЗильк ГмбХ | Способ модификации белка шелка пауков, модифицированный белок шелка пауков, кодирующая его последовательность нуклеиновой кислоты, вектор и клетка-хозяин для его экспрессии |
CN101370556A (zh) | 2006-01-20 | 2009-02-18 | 巴斯夫欧洲公司 | 蛋白质微粒在化妆品中的用途 |
WO2007100524A2 (en) | 2006-02-23 | 2007-09-07 | Genencor International, Inc. | Repeat sequence protein polymer nanoparticles optionally containing active agents and their preparation |
US7751985B2 (en) | 2006-12-14 | 2010-07-06 | The Hong Kong Polytechnic University | Process for preparing silk polypeptide in a controllable particle size |
WO2009069123A2 (en) | 2007-11-26 | 2009-06-04 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Compositions comprising fibrous polypeptides and polysaccharides |
EP2328567A2 (de) * | 2008-08-08 | 2011-06-08 | Basf Se | Wirkstoffhaltige fasernflächengebilde auf basis von biopolymeren, ihre anwendungen und verfahren zu ihrer herstellung |
JP5789799B2 (ja) | 2009-08-21 | 2015-10-07 | 国立研究開発法人農業生物資源研究所 | 多孔質体の製造方法、細胞又は組織供給用支持体の製造方法、及び組織供給体の製造方法 |
WO2011063990A2 (en) | 2009-11-30 | 2011-06-03 | Ludwig-Maximilians-Universität München | Silk particles for controlled and sustained delivery of compounds |
EP2560683B2 (en) | 2010-04-23 | 2022-07-20 | F. Hoffmann-La Roche AG | Production of heteromultimeric proteins |
CA2812063A1 (en) | 2010-08-30 | 2012-06-07 | President And Fellows Of Harvard College | Shear controlled release for stenotic lesions and thrombolytic therapies |
JP5704687B2 (ja) | 2010-10-06 | 2015-04-22 | 日立化成株式会社 | シルクフィブロイン多孔質体 |
JP5704688B2 (ja) | 2010-10-06 | 2015-04-22 | 日立化成株式会社 | シルクフィブロイン多孔質体の製造方法 |
WO2012116439A1 (en) | 2011-02-28 | 2012-09-07 | Mcgill University | Biomaterial, method for making the biomaterial and uses of the same |
CN102344686A (zh) | 2011-08-02 | 2012-02-08 | 北京汇亨创管理咨询有限公司 | 一种以聚乙烯醇为稳定剂的丝素蛋白纳米颗粒的制备方法 |
CN103827139B (zh) | 2011-11-02 | 2016-11-09 | 丝芭博株式会社 | 多肽溶液和使用了该多肽溶液的人造多肽纤维的制造方法以及多肽的精制方法 |
RU2478706C1 (ru) * | 2011-12-23 | 2013-04-10 | Игорь Иванович Агапов | Способ получения суспензий гидрогелевых микрочастиц с заданными размерами на основе рекомбинантного белка паутины и их применение |
US9439972B2 (en) | 2012-09-10 | 2016-09-13 | Ad Lunam Labs, Inc. | Antifungal serum |
CN104918950B (zh) | 2012-12-27 | 2018-10-12 | 丝芭博株式会社 | 亲水性重组蛋白的部分精制方法 |
JP6077569B2 (ja) | 2012-12-27 | 2017-02-08 | Spiber株式会社 | 親水性組換えタンパク質の抽出方法 |
CN104936979B (zh) | 2013-04-25 | 2020-02-21 | 丝芭博株式会社 | 多肽颗粒及其制造方法 |
WO2014175178A1 (ja) | 2013-04-25 | 2014-10-30 | スパイバー株式会社 | ポリペプチド多孔質体及びその製造方法 |
-
2014
- 2014-04-18 EP EP14788691.5A patent/EP2990062B1/en active Active
- 2014-04-18 CN CN201480005729.6A patent/CN104936625A/zh active Pending
- 2014-04-18 JP JP2015513725A patent/JP5782580B2/ja active Active
- 2014-04-18 WO PCT/JP2014/061041 patent/WO2014175177A1/ja active Application Filing
- 2014-04-18 US US14/764,455 patent/US9968682B2/en active Active
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020067572A1 (ja) | 2018-09-28 | 2020-04-02 | Spiber株式会社 | タンパク質組成物の製造方法 |
WO2021065794A1 (ja) | 2019-09-30 | 2021-04-08 | Spiber株式会社 | タンパク質成形体の製造方法 |
WO2021066078A1 (ja) | 2019-09-30 | 2021-04-08 | Spiber株式会社 | 筋組織再生剤 |
Also Published As
Publication number | Publication date |
---|---|
US9968682B2 (en) | 2018-05-15 |
EP2990062A4 (en) | 2017-01-04 |
WO2014175177A1 (ja) | 2014-10-30 |
EP2990062A1 (en) | 2016-03-02 |
US20150374833A1 (en) | 2015-12-31 |
EP2990062B1 (en) | 2021-11-10 |
CN104936625A (zh) | 2015-09-23 |
JPWO2014175177A1 (ja) | 2017-02-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5782580B2 (ja) | ポリペプチドヒドロゲル及びその製造方法 | |
JP5796147B2 (ja) | ポリペプチド多孔質体及びその製造方法 | |
JP5823079B2 (ja) | ポリペプチドパーティクルの製造方法 | |
JP5427322B2 (ja) | ポリペプチドの溶液とこれを用いた人造ポリペプチド繊維の製造方法及びポリペプチドの精製方法 | |
JP7088511B2 (ja) | フィブロイン様タンパク質を含むコンポジット成形組成物及びその製造方法 | |
WO2013065651A1 (ja) | タンパク質溶液及びこれを用いたタンパク質繊維の製造方法 | |
EP2940033B1 (en) | Partial purification method for hydrophilic recombinant protein | |
JP6856828B2 (ja) | 極性溶媒溶液及びその製造方法 | |
WO2017094722A1 (ja) | タンパク質溶液を製造する方法 | |
US11668024B2 (en) | Polar solvent solution and production method thereof | |
JP7270978B2 (ja) | ポリペプチド溶液、及びポリペプチド繊維の製造方法、並びに人造ポリペプチド | |
WO2019194231A1 (ja) | タンパク質組成物及びその製造方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20150624 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20150707 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20150717 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5782580 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S303 | Written request for registration of pledge or change of pledge |
Free format text: JAPANESE INTERMEDIATE CODE: R316303 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S321 | Written request for registration of change in pledge agreement |
Free format text: JAPANESE INTERMEDIATE CODE: R316321 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
S303 | Written request for registration of pledge or change of pledge |
Free format text: JAPANESE INTERMEDIATE CODE: R316303 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
S321 | Written request for registration of change in pledge agreement |
Free format text: JAPANESE INTERMEDIATE CODE: R316321 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |