JP4430677B2 - B型肝炎ウイルスs抗原の検出法 - Google Patents
B型肝炎ウイルスs抗原の検出法 Download PDFInfo
- Publication number
- JP4430677B2 JP4430677B2 JP2006536401A JP2006536401A JP4430677B2 JP 4430677 B2 JP4430677 B2 JP 4430677B2 JP 2006536401 A JP2006536401 A JP 2006536401A JP 2006536401 A JP2006536401 A JP 2006536401A JP 4430677 B2 JP4430677 B2 JP 4430677B2
- Authority
- JP
- Japan
- Prior art keywords
- antigen
- virus
- hepatitis
- antibody
- hbs
- 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
- 239000000427 antigen Substances 0.000 title claims description 194
- 108091007433 antigens Proteins 0.000 title claims description 194
- 102000036639 antigens Human genes 0.000 title claims description 194
- 241000700721 Hepatitis B virus Species 0.000 title claims description 77
- 238000000034 method Methods 0.000 title claims description 48
- 239000003795 chemical substances by application Substances 0.000 claims description 45
- 239000004094 surface-active agent Substances 0.000 claims description 45
- 108090000623 proteins and genes Proteins 0.000 claims description 35
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 29
- 102000004169 proteins and genes Human genes 0.000 claims description 27
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 22
- 238000001514 detection method Methods 0.000 claims description 19
- 239000002535 acidifier Substances 0.000 claims description 15
- 239000003398 denaturant Substances 0.000 claims description 13
- 239000003638 chemical reducing agent Substances 0.000 claims description 9
- 241000700605 Viruses Species 0.000 claims description 7
- 238000009739 binding Methods 0.000 claims description 6
- 239000003153 chemical reaction reagent Substances 0.000 claims description 6
- 208000002672 hepatitis B Diseases 0.000 claims description 3
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 3
- 238000010494 dissociation reaction Methods 0.000 claims 2
- 230000005593 dissociations Effects 0.000 claims 2
- 239000000523 sample Substances 0.000 description 83
- 239000000243 solution Substances 0.000 description 43
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 40
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 30
- 235000018102 proteins Nutrition 0.000 description 22
- 239000000232 Lipid Bilayer Substances 0.000 description 21
- 230000000890 antigenic effect Effects 0.000 description 21
- 238000005259 measurement Methods 0.000 description 21
- 239000002245 particle Substances 0.000 description 21
- 239000011780 sodium chloride Substances 0.000 description 20
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 18
- 210000004408 hybridoma Anatomy 0.000 description 15
- 238000005406 washing Methods 0.000 description 14
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 13
- 150000001413 amino acids Chemical class 0.000 description 13
- 239000004202 carbamide Substances 0.000 description 13
- 230000000694 effects Effects 0.000 description 13
- 208000015181 infectious disease Diseases 0.000 description 12
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 230000035945 sensitivity Effects 0.000 description 10
- 239000012528 membrane Substances 0.000 description 9
- 210000004379 membrane Anatomy 0.000 description 9
- 238000002156 mixing Methods 0.000 description 9
- 239000002244 precipitate Substances 0.000 description 9
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 8
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 8
- 229960000789 guanidine hydrochloride Drugs 0.000 description 8
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 239000012798 spherical particle Substances 0.000 description 8
- 230000000903 blocking effect Effects 0.000 description 7
- 238000005119 centrifugation Methods 0.000 description 7
- 239000012634 fragment Substances 0.000 description 7
- 230000035772 mutation Effects 0.000 description 7
- 102000013415 peroxidase activity proteins Human genes 0.000 description 7
- 108040007629 peroxidase activity proteins Proteins 0.000 description 7
- 239000008363 phosphate buffer Substances 0.000 description 7
- 230000009257 reactivity Effects 0.000 description 7
- 239000012064 sodium phosphate buffer Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 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 6
- 241000588724 Escherichia coli Species 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 6
- 239000003513 alkali Substances 0.000 description 6
- 125000000539 amino acid group Chemical group 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 6
- 238000000691 measurement method Methods 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 5
- 239000002671 adjuvant Substances 0.000 description 5
- 239000003945 anionic surfactant Substances 0.000 description 5
- 239000003093 cationic surfactant Substances 0.000 description 5
- 239000013613 expression plasmid Substances 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 108091008146 restriction endonucleases Proteins 0.000 description 5
- 239000011734 sodium Substances 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 4
- 102000011632 Caseins Human genes 0.000 description 4
- 108010076119 Caseins Proteins 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- 229920001213 Polysorbate 20 Polymers 0.000 description 4
- 239000012980 RPMI-1640 medium Substances 0.000 description 4
- 125000000217 alkyl group Chemical group 0.000 description 4
- 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 4
- 229960000723 ampicillin Drugs 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 230000003053 immunization Effects 0.000 description 4
- 229910001629 magnesium chloride Inorganic materials 0.000 description 4
- 230000003472 neutralizing effect Effects 0.000 description 4
- 239000002736 nonionic surfactant Substances 0.000 description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 4
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 150000003512 tertiary amines Chemical class 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 3
- OGMADIBCHLQMIP-UHFFFAOYSA-N 2-aminoethanethiol;hydron;chloride Chemical compound Cl.NCCS OGMADIBCHLQMIP-UHFFFAOYSA-N 0.000 description 3
- 206010003445 Ascites Diseases 0.000 description 3
- 101710132601 Capsid protein Proteins 0.000 description 3
- 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 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- 101150010882 S gene Proteins 0.000 description 3
- 239000002280 amphoteric surfactant Substances 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 230000002163 immunogen Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 229940080237 sodium caseinate Drugs 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- CMBJWSUWPPPZTH-UHFFFAOYSA-N 1-(diethylamino)ethanethiol;hydrochloride Chemical compound Cl.CCN(CC)C(C)S CMBJWSUWPPPZTH-UHFFFAOYSA-N 0.000 description 2
- UMCMPZBLKLEWAF-BCTGSCMUSA-N 3-[(3-cholamidopropyl)dimethylammonio]propane-1-sulfonate Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCC[N+](C)(C)CCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 UMCMPZBLKLEWAF-BCTGSCMUSA-N 0.000 description 2
- PBVAJRFEEOIAGW-UHFFFAOYSA-N 3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochloride Chemical compound Cl.OC(=O)CCP(CCC(O)=O)CCC(O)=O PBVAJRFEEOIAGW-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 108010017384 Blood Proteins Proteins 0.000 description 2
- 102000004506 Blood Proteins Human genes 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241001131785 Escherichia coli HB101 Species 0.000 description 2
- 206010019799 Hepatitis viral Diseases 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- BACYUWVYYTXETD-UHFFFAOYSA-N N-Lauroylsarcosine Chemical compound CCCCCCCCCCCC(=O)N(C)CC(O)=O BACYUWVYYTXETD-UHFFFAOYSA-N 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 238000001042 affinity chromatography Methods 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000003113 dilution method Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 231100000283 hepatitis Toxicity 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 208000019423 liver disease Diseases 0.000 description 2
- 201000000050 myeloid neoplasm Diseases 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 210000002381 plasma Anatomy 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- -1 polypropylene Polymers 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- XOJVVFBFDXDTEG-UHFFFAOYSA-N pristane Chemical compound CC(C)CCCC(C)CCCC(C)CCCC(C)C XOJVVFBFDXDTEG-UHFFFAOYSA-N 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000011535 reaction buffer Substances 0.000 description 2
- 108700004121 sarkosyl Proteins 0.000 description 2
- 150000003431 steroids Chemical group 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 201000001862 viral hepatitis Diseases 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 239000002888 zwitterionic surfactant Substances 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- SXOUIMVOMIGLHO-AATRIKPKSA-N (E)-3-(indol-2-yl)acrylic acid Chemical compound C1=CC=C2NC(/C=C/C(=O)O)=CC2=C1 SXOUIMVOMIGLHO-AATRIKPKSA-N 0.000 description 1
- 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 1
- TVZGACDUOSZQKY-LBPRGKRZSA-N 4-aminofolic acid Chemical compound C1=NC2=NC(N)=NC(N)=C2N=C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 TVZGACDUOSZQKY-LBPRGKRZSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 101001094887 Ambrosia artemisiifolia Pectate lyase 1 Proteins 0.000 description 1
- 101001123576 Ambrosia artemisiifolia Pectate lyase 2 Proteins 0.000 description 1
- 101001123572 Ambrosia artemisiifolia Pectate lyase 3 Proteins 0.000 description 1
- 101000573177 Ambrosia artemisiifolia Pectate lyase 5 Proteins 0.000 description 1
- 101000942941 Arabidopsis thaliana DNA ligase 6 Proteins 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 101100283604 Caenorhabditis elegans pigk-1 gene Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 101710094648 Coat protein Proteins 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 241000233866 Fungi 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
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 description 1
- 101000872838 Hepatitis B virus genotype C subtype adr (isolate China/NC-1/1988) Small envelope protein Proteins 0.000 description 1
- 241000724675 Hepatitis E virus Species 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 101710084021 Large envelope protein Proteins 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 101710125418 Major capsid protein Proteins 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 125000000729 N-terminal amino-acid group Chemical group 0.000 description 1
- 206010029719 Nonspecific reaction Diseases 0.000 description 1
- 101710141454 Nucleoprotein Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 208000037581 Persistent Infection Diseases 0.000 description 1
- 101001006139 Podospora anserina Heterokaryon incompatibility protein s Proteins 0.000 description 1
- 241000276498 Pollachius virens Species 0.000 description 1
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 101710083689 Probable capsid protein Proteins 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 239000012506 Sephacryl® Substances 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 101000980463 Treponema pallidum (strain Nichols) Chaperonin GroEL Proteins 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 108700005077 Viral Genes Proteins 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 230000003113 alkalizing effect Effects 0.000 description 1
- 229960003896 aminopterin Drugs 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000003196 chaotropic effect Effects 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000007771 core particle Substances 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- YBDSNEVSFQMCTL-UHFFFAOYSA-O diethyl(2-sulfanylethyl)azanium Chemical compound CC[NH+](CC)CCS YBDSNEVSFQMCTL-UHFFFAOYSA-O 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- SFNALCNOMXIBKG-UHFFFAOYSA-N ethylene glycol monododecyl ether Chemical compound CCCCCCCCCCCCOCCO SFNALCNOMXIBKG-UHFFFAOYSA-N 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 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
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002736 metal compounds Chemical class 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 108091005601 modified peptides Proteins 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 239000000941 radioactive substance Substances 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 230000015227 regulation of liquid surface tension Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
- C07K14/01—DNA viruses
- C07K14/02—Hepadnaviridae, e.g. hepatitis B virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
- C07K16/082—Hepadnaviridae, e.g. hepatitis B virus
-
- 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
- C12N15/02—Preparation of hybrid cells by fusion of two or more cells, e.g. protoplast fusion
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
-
- 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
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Urology & Nephrology (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Communicable Diseases (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Zoology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Plant Pathology (AREA)
- Peptides Or Proteins (AREA)
Description
Howard et al. Viral Hepatitis and Liver Disease (ed by Zuckerman AJ, Alan R), p1094−1101, Liss Inc, New York, 1988。 岡本宏明 日本臨床 分子肝炎ウイルス病学 基礎・臨床・予防 下巻 A,B,D,E型肝炎ウイルス編、p212−222、平成7年10月26日発行。 Thiers et al.Lancet, ii, 1273−1276, 1988。
以下の実施例は本発明を例証するものであるが、これによって本発明の範囲を制限するものではない。
(A)TrpE−HBs(26〜80)抗原発現プラスミドの構築
HBs(26〜80)領域に相当する発現プラスミドは以下の方法で構築した。HBV患者血清100μlをDNA抽出液100μl[1M Tris−HCl(pH8.4)が10μl,250mM EDTAが8μl,10%SDSが40μl,5M NaClが8μl,20mg/ml ProteinaseKが10μl,tRNA(5μg/μl)が1μl,滅菌水が23μl]と混合させ、54℃、30分間保温した。200μlのフェノール・クロロホルム(1:1)溶液を加えて混合し、15Krpmで5分間の遠心分離の後、上清を取り出し150μlのイソプロパノールと7μlの5M NaClを加えて−20℃1時間静置した。15Krpm、4℃で5分間遠心分離し、沈殿物を70%エタノールでリンスし、15Krpm、4℃で再度5分間遠心分離した。沈殿物を自然乾燥させ、20μlの滅菌水に溶解させてHBV DNA溶液とした。
発現プラスミドpATtrpE−HBs(26〜80)をもつ大腸菌HB101株を50μg/mlのアンピシリンを含む3mlの2YT培地(1.6%トリプトン、1%酵母エキス、0.5%NaCl)に接種し、37℃で9時間培養する。この培養液1mlを50μg/mlのアンピシリンを含む100mlのM9−CA培地(0.6%NA2 HPO4 ,0.5%KH2 PO4 ,0.5%NaCl,0.1%NH4Cl,0.1mM CaCl2,2mM MgSO4,0.5%カザミノ酸、0.2%グルコース)に植え継ぎ、37℃で培養した。OD600=0.3の時に終濃度40mg/lになるようにインドールアクリル酸を加え、さらに16時間培養した。この培養液を5Krpm、10分間遠心分離して菌体を集めた。
前記方法により調製したポリペプチド[trpE−HBs(26〜80)]を6M尿素溶解後、0.15M NaClを含む10mMリン酸緩衝液(pH7.3)(PBS)に終濃度が0.2〜1.0mg/mlとなるように希釈し、等量のフロイントアジュバントと混和し、4〜6週令のBALB/cマウスに10−20μg腹腔内投与した。2−4週間ごとに同様の追加免疫を行い、さらにPBSに溶解したHBs 10μgを最終免疫として尾静脈内に投与した。
実施例2に記載の方法により得られたハイブリドーマを、あらかじめプリスタンを腹腔投与しておいたBALB/cマウス腹腔に移植し、腹水中に産生されてくるモノクローナル抗体を取得した。該モノクローナル抗体は、プロテインAセファロースカラムを用いたアフィニティークロマトグラフィーによりIgGフラクションを分離精製した。
抗HBs抗原モノクローナル抗体6G6を終濃度が6μg/mlになるように0.15M NaClを含む10mMリン酸ナトリウム緩衝液(pH7.3)で希釈し、96ウエルマイクロプレート(ヌンク社製)1ウエルにつき80μlづつ分注した。4℃で一晩静置後、0.15M NaClを含む10mMリン酸ナトリウム緩衝液(pH7.3)0.35mlを用いて2回洗浄し、0.5%カゼイン−Naを含む10mMリン酸ナトリウム緩衝液(pH7.3)(以下ブロッキング液)、0.35mlを添加し、さらに室温で2時間静置した。
HBs抗原陰性であるが、HBVに感染していると考えられる検体を、実施例4の方法を改良した方法で測定した。
HBV抗原陰性検体または抗HBs抗体を含むHBV抗原陽性検体(#990493、#990640、#990650)50μLに、各種濃度の塩酸水溶液50μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、以下に記す測定法を用いて検討した。
HBV抗原陰性検体またはHBs抗原陽性検体(#990493、#990640、#990650)30μLに、各種界面活性剤を1.0N塩酸濃度の水溶液に溶解したその30μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、1)で前述した方法で検討した(表6〜表9)。尚、表に示した塩酸濃度及び界面活性剤濃度は、検体(Sample)と処理剤を混合後の処理時濃度で表した。
HBV陰性検体またはHBs抗原陽性検体(#990493、#990640、#990650)30μLに、蛋白質変性剤である尿素またはグアニジン塩酸塩を1.0N塩酸濃度の水溶液に溶解し、その30μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、1)で前述した方法で検討した。各々のHBs抗原陽性検体の免疫活性を表10に示した。尚、表10に示した塩酸濃度及び蛋白質変性剤濃度は、検体(Sample)と処理剤を混合後の処理時濃度で表した。
HBV抗原陰性検体(Normal plasma)または3例のHBs抗原陽性検体(#990493、#990640、#990650)30μLに、1.0N塩酸を含む溶液に還元剤であるジチオスレイトール、2−メルカプトエチルアミン塩酸塩、2−ジエチルアミノエタンチオール塩酸塩を混合した溶液の30μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、1)で述べた方法を用いて検討した(表11)。
HBV抗原陰性検体または抗HBs抗体を含むHBV抗原陽性検体(#990493、#990640、#990650)50μLに、各種濃度の水酸化ナトリウム水溶液50μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、以下に記す測定法を用いて検討した。
HBV抗原陰性検体またはHBs抗原陽性検体(#990493、#990640、#990650)30μLに、各種界面活性剤を1.0N水酸化ナトリウム濃度の水溶液に溶解したその30μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、5)で前述した方法で検討した(表13〜表17)。尚、表に示した水酸化ナトリウム濃度及び界面活性剤濃度は、検体(Sample)と処理剤を混合後の処理時濃度で表した。
HBs抗原陰性検体またはHBs抗原陽性検体(#990493、#990640、#990650)30μLに、蛋白質変性剤である尿素またはグアニジン塩酸塩を1.0N水酸化ナトリウム濃度の水溶液に溶解し、その30μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、5)で前述した方法で検討した。各々のHBs抗原陽性検体の免疫活性を表18に示した。尚、表18に示した水酸化ナトリウム濃度及び蛋白質変性剤濃度は、検体(Sample)と処理剤を混合後の処理時濃度で表した。
HBV抗原陰性検体または3例のHBs抗原陽性検体(#990493、#990640、#990650)30μLに、1.0N水酸化ナトリウムを含む溶液に還元剤であるジチオスレイトール、2−メルカプトエチルアミン塩酸塩、ジエチルアミノエタンチオール塩酸塩、2−メルカプトエタノール、トリ(2−カルボキシエチル)ホスフィン塩酸塩を混合した溶液の30μLを添加して、室温で10分間インキュベーションを行い、その50μLを測定試料として、5)で述べた方法を用いて検討した(表19)。
Claims (12)
- B型肝炎ウイルスs抗原の26番目から80番目に相当するアミノ酸配列からなるペプチドを認識する抗体。
- 配列番号1のアミノ酸配列からなるペプチドを認識する抗体。
- 変性剤で変性させたB型肝炎ウイルスs抗原の26番目から80番目に相当するアミノ酸配列からなるまたは配列番号1に示されたアミノ酸配列からなるペプチドを抗原として得られる、B型肝炎ウイルスs抗原を認識することのできる請求項1または2に記載の抗体。
- 請求項1〜3の何れかに記載の抗体を用いる、B型肝炎ウイルスまたはB型肝炎ウイルスs抗原の検出法。
- 変性剤の共存下で行う、請求項4に記載の検出法。
- 変性剤が界面活性剤である、請求項4または5に記載の検出法。
- 変性剤が(1)酸性化剤、および(2)界面活性剤および/または蛋白質変性剤を含有する処理剤で処理することにより、B型肝炎ウイルスs抗原の解離とB型肝炎ウイルスs抗原に結合する抗体の不活化を行うことを特徴とする請求項4または5に記載の検出法。
- 変性剤が(1)アルカリ剤、および(2)界面活性剤および/または蛋白質変性剤を含有する処理剤で処理することにより、B型肝炎ウイルスs抗原の解離とB型肝炎ウイルスs抗原に結合する抗体の不活化を行うことを特徴とする請求項4または5に記載の検出法。
- 変性剤が(1)アルカリ剤、および(2)界面活性剤および/または蛋白質変性剤および/または還元剤を含有する処理剤で処理することにより、B型肝炎ウイルスs抗原の解離とB型肝炎ウイルスs抗原に結合する抗体の不活化を行うことを特徴とする請求項4または5に記載の検出法。
- さらにB型肝炎ウイルスs抗原以外のB型肝炎ウイルス抗原に対する抗体を利用する、請求項4〜6の何れかに記載の検出法。
- 請求項1〜3の何れかに記載の抗体および変性剤を含む、B型肝炎ウイルスs抗原検出用試薬。
- 請求項1〜3の何れかに記載の抗体、B型肝炎ウイルスs抗原以外のB型肝炎ウイルス抗原に対する抗体および変性剤を含む、B型肝炎ウイルスs抗原検出用試薬。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004274852 | 2004-09-22 | ||
JP2004274852 | 2004-09-22 | ||
PCT/JP2005/017420 WO2006033368A1 (ja) | 2004-09-22 | 2005-09-21 | B型肝炎ウイルスs抗原の検出法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2006033368A1 JPWO2006033368A1 (ja) | 2008-05-15 |
JP4430677B2 true JP4430677B2 (ja) | 2010-03-10 |
Family
ID=36090125
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2006536401A Active JP4430677B2 (ja) | 2004-09-22 | 2005-09-21 | B型肝炎ウイルスs抗原の検出法 |
Country Status (8)
Country | Link |
---|---|
US (1) | US8679762B2 (ja) |
EP (1) | EP1806363B1 (ja) |
JP (1) | JP4430677B2 (ja) |
KR (1) | KR100916992B1 (ja) |
CN (1) | CN101023098B (ja) |
CA (1) | CA2580620C (ja) |
ES (1) | ES2428630T3 (ja) |
WO (1) | WO2006033368A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019107279A1 (ja) | 2017-11-30 | 2019-06-06 | 富士レビオ株式会社 | B型肝炎ウイルスs抗原の測定方法及び測定キット |
Families Citing this family (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP4855126B2 (ja) * | 2006-04-07 | 2012-01-18 | アボットジャパン株式会社 | パルボウイルスb19抗原測定方法 |
CA2667859C (en) | 2006-10-30 | 2017-01-24 | Advanced Life Science Institute, Inc. | Method for analysis of hepatitis b virus s antigen |
JP5332011B2 (ja) * | 2006-10-30 | 2013-11-06 | 株式会社先端生命科学研究所 | B型肝炎ウイルスの高感度免疫学的分析方法及び免疫学的分析用試薬 |
JP5351724B2 (ja) * | 2009-11-30 | 2013-11-27 | シスメックス株式会社 | C型肝炎ウイルスのコア蛋白の検出方法及び検出用試薬キット |
CN102094002A (zh) * | 2009-12-11 | 2011-06-15 | 上海裕隆临床检验中心有限公司 | 乙型肝炎病毒dna提取试剂及提取方法 |
CN102183647A (zh) * | 2011-01-24 | 2011-09-14 | 杭州师范大学 | 一种乙型肝炎病毒表面抗原的检测试剂及检测方法 |
EP2950099B1 (en) * | 2013-01-28 | 2018-09-19 | Sysmex Corporation | Pretreatment method for sample for detecting hbs antigen, and use therefor |
US10634676B2 (en) * | 2015-10-15 | 2020-04-28 | Fujirebio Inc. | Method and kit for simultaneously detecting human parvovirus B19 antigen and antibody |
MX2020002939A (es) | 2017-09-18 | 2020-07-22 | Bayer Healthcare Llc | Metodos de inactivacion de virus utilizando n-metilglutamida y sus derivados. |
CN109870581B (zh) * | 2017-12-04 | 2021-05-04 | 厦门万泰凯瑞生物技术有限公司 | 一种定量检测HBsAg的试剂盒及方法 |
CN109870571A (zh) * | 2018-12-29 | 2019-06-11 | 广东云天抗体生物科技有限公司 | 一种检测猴g-csf的酶联免疫试剂盒 |
CN109870570A (zh) * | 2018-12-29 | 2019-06-11 | 广东云天抗体生物科技有限公司 | 一种检测猴il-18的酶联免疫试剂盒 |
CN115710299B (zh) * | 2022-10-31 | 2024-05-24 | 北京工业大学 | 肝靶向的早期药物性肝炎和自身免疫性肝炎的荧光/光声双模态探针 |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4861588A (en) * | 1985-02-05 | 1989-08-29 | New York Blood Center, Inc. | Pre-S gene coded peptide hepatitis B immunogens, vaccines, diagnostics, and synthetic lipid vesicle carriers |
JPH089637B2 (ja) * | 1986-06-18 | 1996-01-31 | 財団法人阪大微生物病研究会 | B型肝炎ウイルス抗原とその製造法 |
GB9007024D0 (en) | 1990-03-29 | 1990-05-30 | Imperial College | Novel vaccine |
CN1121357A (zh) | 1993-03-24 | 1996-04-24 | 科技医药皇家学院 | 乙型肝炎逃逸突变体特异性结合分子 |
US5531990A (en) | 1993-12-15 | 1996-07-02 | Health Research, Inc. | Method of raising an immune response with an anti-idiotypic antibody having correspondence with human hepatitis B surface antigen |
GB9401987D0 (en) | 1994-02-02 | 1994-03-30 | Imperial College | Modified nucleic acid |
EP0919568A1 (en) | 1997-12-01 | 1999-06-02 | Sorin Diagnostics S.r.l. | Escape mutant of the surface antigen of hepatitis B virus |
AU756858B2 (en) | 1998-06-19 | 2003-01-23 | Government Of The Republic Of Singapore | A vaccine-induced hepatitis B viral strain and uses thereof |
JP3847257B2 (ja) | 2000-08-11 | 2006-11-22 | 株式会社先端生命科学研究所 | Hbvの検出又は測定方法 |
-
2005
- 2005-09-21 JP JP2006536401A patent/JP4430677B2/ja active Active
- 2005-09-21 CA CA2580620A patent/CA2580620C/en active Active
- 2005-09-21 EP EP05785966.2A patent/EP1806363B1/en active Active
- 2005-09-21 ES ES05785966T patent/ES2428630T3/es active Active
- 2005-09-21 KR KR1020077006465A patent/KR100916992B1/ko active IP Right Grant
- 2005-09-21 US US11/663,517 patent/US8679762B2/en active Active
- 2005-09-21 CN CN2005800314440A patent/CN101023098B/zh active Active
- 2005-09-21 WO PCT/JP2005/017420 patent/WO2006033368A1/ja active Application Filing
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019107279A1 (ja) | 2017-11-30 | 2019-06-06 | 富士レビオ株式会社 | B型肝炎ウイルスs抗原の測定方法及び測定キット |
JPWO2019107279A1 (ja) * | 2017-11-30 | 2020-12-24 | 富士レビオ株式会社 | B型肝炎ウイルスs抗原の測定方法及び測定キット |
US11454632B2 (en) | 2017-11-30 | 2022-09-27 | Fujirebio Inc. | Assay method and assay kit for hepatitis B virus S antigen |
JP7198222B2 (ja) | 2017-11-30 | 2022-12-28 | 富士レビオ株式会社 | B型肝炎ウイルスs抗原の測定方法及び測定キット |
Also Published As
Publication number | Publication date |
---|---|
JPWO2006033368A1 (ja) | 2008-05-15 |
WO2006033368A1 (ja) | 2006-03-30 |
KR100916992B1 (ko) | 2009-09-14 |
CN101023098B (zh) | 2012-07-18 |
CA2580620A1 (en) | 2006-03-30 |
EP1806363A4 (en) | 2007-12-12 |
US20080193916A1 (en) | 2008-08-14 |
EP1806363A1 (en) | 2007-07-11 |
KR20070056107A (ko) | 2007-05-31 |
US8679762B2 (en) | 2014-03-25 |
CN101023098A (zh) | 2007-08-22 |
ES2428630T3 (es) | 2013-11-08 |
EP1806363B1 (en) | 2013-09-04 |
CA2580620C (en) | 2014-04-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP4430677B2 (ja) | B型肝炎ウイルスs抗原の検出法 | |
KR100847586B1 (ko) | C형 간염 바이러스의 측정방법 | |
US8546075B2 (en) | Method of detecting hepatitis C virus | |
US20080138794A1 (en) | Method for detecting or measuring HBV | |
JP3217600B2 (ja) | 非a非b型肝炎ウイルス関連抗原のイムノアッセイ、それに使用するモノクローナル抗体、およびこの抗体を産生するハイブリドーマ | |
EP1724285B1 (en) | Anti-HBs monoclonal antibodies | |
JP2004500041A (ja) | 新規なhev抗原性ペプチド及び方法 | |
JP5261822B2 (ja) | B型肝炎ウイルスs抗原の分析方法 | |
US20040073006A1 (en) | Antibody against norwalk virus and method of detecting virus by using the antibody | |
US7105165B2 (en) | Mutant human hepatitis B viral strain and uses thereof | |
JP3176570B2 (ja) | Hcvの検出又は測定方法 | |
KR20240009996A (ko) | 항노로바이러스 항체 | |
KR20240009995A (ko) | 항노로바이러스 항체 | |
KR20240009997A (ko) | 노로바이러스의 면역 측정 방법 및 면역 측정 기구 | |
JP2001224371A (ja) | Hcvの検出又は測定方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20090811 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20091009 |
|
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: 20091201 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20091217 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20121225 Year of fee payment: 3 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 4430677 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20121225 Year of fee payment: 3 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20121225 Year of fee payment: 3 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20131225 Year of fee payment: 4 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S531 | Written request for registration of change of domicile |
Free format text: JAPANESE INTERMEDIATE CODE: R313531 |
|
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 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |