JPS6223825B2 - - Google Patents
Info
- Publication number
- JPS6223825B2 JPS6223825B2 JP55120595A JP12059580A JPS6223825B2 JP S6223825 B2 JPS6223825 B2 JP S6223825B2 JP 55120595 A JP55120595 A JP 55120595A JP 12059580 A JP12059580 A JP 12059580A JP S6223825 B2 JPS6223825 B2 JP S6223825B2
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- dye
- antigen
- labeled
- labeled antigen
- 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
Links
- -1 silver halide Chemical class 0.000 claims description 23
- 239000000427 antigen Substances 0.000 claims description 16
- 102000036639 antigens Human genes 0.000 claims description 16
- 108091007433 antigens Proteins 0.000 claims description 16
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 16
- 230000003595 spectral effect Effects 0.000 claims description 11
- 229910052709 silver Inorganic materials 0.000 claims description 10
- 239000004332 silver Substances 0.000 claims description 10
- 230000001235 sensitizing effect Effects 0.000 claims description 9
- 238000002372 labelling Methods 0.000 claims description 7
- 239000000126 substance Substances 0.000 claims description 7
- DZVCFNFOPIZQKX-LTHRDKTGSA-M merocyanine Chemical compound [Na+].O=C1N(CCCC)C(=O)N(CCCC)C(=O)C1=C\C=C\C=C/1N(CCCS([O-])(=O)=O)C2=CC=CC=C2O\1 DZVCFNFOPIZQKX-LTHRDKTGSA-M 0.000 claims description 5
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims description 4
- 239000007795 chemical reaction product Substances 0.000 claims description 4
- 125000000623 heterocyclic group Chemical group 0.000 claims description 3
- 230000036046 immunoreaction Effects 0.000 claims description 3
- 125000001424 substituent group Chemical group 0.000 claims description 3
- 238000003018 immunoassay Methods 0.000 claims description 2
- 230000003287 optical effect Effects 0.000 claims description 2
- 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 27
- 239000000975 dye Substances 0.000 description 22
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 15
- 102000004877 Insulin Human genes 0.000 description 11
- 108090001061 Insulin Proteins 0.000 description 11
- 229940125396 insulin Drugs 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 238000000034 method Methods 0.000 description 9
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 108010005991 Pork Regular Insulin Proteins 0.000 description 5
- 125000003118 aryl group Chemical group 0.000 description 5
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 101001018100 Homo sapiens Lysozyme C Proteins 0.000 description 4
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 3
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 125000000217 alkyl group Chemical group 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- WFIYPADYPQQLNN-UHFFFAOYSA-N 2-[2-(4-bromopyrazol-1-yl)ethyl]isoindole-1,3-dione Chemical compound C1=C(Br)C=NN1CCN1C(=O)C2=CC=CC=C2C1=O WFIYPADYPQQLNN-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
- YOETUEMZNOLGDB-UHFFFAOYSA-N 2-methylpropyl carbonochloridate Chemical compound CC(C)COC(Cl)=O YOETUEMZNOLGDB-UHFFFAOYSA-N 0.000 description 2
- QMHIMXFNBOYPND-UHFFFAOYSA-N 4-methylthiazole Chemical compound CC1=CSC=N1 QMHIMXFNBOYPND-UHFFFAOYSA-N 0.000 description 2
- AAKPXIJKSNGOCO-UHFFFAOYSA-N 5-phenyl-1,3-benzothiazole Chemical compound C=1C=C2SC=NC2=CC=1C1=CC=CC=C1 AAKPXIJKSNGOCO-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 241000700199 Cavia porcellus Species 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- 125000003107 substituted aryl group Chemical group 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- UUJOCRCAIOAPFK-UHFFFAOYSA-N 1,3-benzoselenazol-5-ol Chemical compound OC1=CC=C2[se]C=NC2=C1 UUJOCRCAIOAPFK-UHFFFAOYSA-N 0.000 description 1
- AIGNCQCMONAWOL-UHFFFAOYSA-N 1,3-benzoselenazole Chemical compound C1=CC=C2[se]C=NC2=C1 AIGNCQCMONAWOL-UHFFFAOYSA-N 0.000 description 1
- FOKHITNCOPPDER-UHFFFAOYSA-N 1,3-benzothiazole;4-chloro-1,3-benzothiazole Chemical compound C1=CC=C2SC=NC2=C1.ClC1=CC=CC2=C1N=CS2 FOKHITNCOPPDER-UHFFFAOYSA-N 0.000 description 1
- UPPYOQWUJKAFSG-UHFFFAOYSA-N 1,3-benzoxazol-5-ol Chemical compound OC1=CC=C2OC=NC2=C1 UPPYOQWUJKAFSG-UHFFFAOYSA-N 0.000 description 1
- SAHAKBXWZLDNAA-UHFFFAOYSA-N 1,3-benzoxazol-6-ol Chemical compound OC1=CC=C2N=COC2=C1 SAHAKBXWZLDNAA-UHFFFAOYSA-N 0.000 description 1
- BCMCBBGGLRIHSE-UHFFFAOYSA-N 1,3-benzoxazole Chemical compound C1=CC=C2OC=NC2=C1 BCMCBBGGLRIHSE-UHFFFAOYSA-N 0.000 description 1
- ALUQMCBDQKDRAK-UHFFFAOYSA-N 2,3,3a,4-tetrahydro-1,3-benzothiazole Chemical compound C1C=CC=C2SCNC21 ALUQMCBDQKDRAK-UHFFFAOYSA-N 0.000 description 1
- 125000000143 2-carboxyethyl group Chemical group [H]OC(=O)C([H])([H])C([H])([H])* 0.000 description 1
- 125000000954 2-hydroxyethyl group Chemical group [H]C([*])([H])C([H])([H])O[H] 0.000 description 1
- 125000004200 2-methoxyethyl group Chemical group [H]C([H])([H])OC([H])([H])C([H])([H])* 0.000 description 1
- QOXOZONBQWIKDA-UHFFFAOYSA-N 3-hydroxypropyl Chemical group [CH2]CCO QOXOZONBQWIKDA-UHFFFAOYSA-N 0.000 description 1
- UWSONZCNXUSTKW-UHFFFAOYSA-N 4,5-Dimethylthiazole Chemical compound CC=1N=CSC=1C UWSONZCNXUSTKW-UHFFFAOYSA-N 0.000 description 1
- BGTVICKPWACXLR-UHFFFAOYSA-N 4,5-diphenyl-1,3-thiazole Chemical compound S1C=NC(C=2C=CC=CC=2)=C1C1=CC=CC=C1 BGTVICKPWACXLR-UHFFFAOYSA-N 0.000 description 1
- NDUHYERSZLRFNL-UHFFFAOYSA-N 4,6-dimethyl-1,3-benzoxazole Chemical compound CC1=CC(C)=C2N=COC2=C1 NDUHYERSZLRFNL-UHFFFAOYSA-N 0.000 description 1
- GQPBBURQQRLAKF-UHFFFAOYSA-N 4-ethyl-1,3-oxazole Chemical compound CCC1=COC=N1 GQPBBURQQRLAKF-UHFFFAOYSA-N 0.000 description 1
- PIUXNZAIHQAHBY-UHFFFAOYSA-N 4-methyl-1,3-benzothiazole Chemical compound CC1=CC=CC2=C1N=CS2 PIUXNZAIHQAHBY-UHFFFAOYSA-N 0.000 description 1
- PUMREIFKTMLCAF-UHFFFAOYSA-N 4-methyl-1,3-oxazole Chemical compound CC1=COC=N1 PUMREIFKTMLCAF-UHFFFAOYSA-N 0.000 description 1
- BJATXNRFAXUVCU-UHFFFAOYSA-N 4-methyl-1,3-selenazole Chemical compound CC1=C[se]C=N1 BJATXNRFAXUVCU-UHFFFAOYSA-N 0.000 description 1
- ZVNPWFOVUDMGRP-UHFFFAOYSA-N 4-methylaminophenol sulfate Chemical compound OS(O)(=O)=O.CNC1=CC=C(O)C=C1.CNC1=CC=C(O)C=C1 ZVNPWFOVUDMGRP-UHFFFAOYSA-N 0.000 description 1
- GHAFJOZKMUPGRQ-UHFFFAOYSA-N 4-nitro-1,3-oxazole Chemical compound [O-][N+](=O)C1=COC=N1 GHAFJOZKMUPGRQ-UHFFFAOYSA-N 0.000 description 1
- HLCQHHLQESOBFS-UHFFFAOYSA-N 4-nitro-1,3-selenazole Chemical compound [O-][N+](=O)C1=C[se]C=N1 HLCQHHLQESOBFS-UHFFFAOYSA-N 0.000 description 1
- RILRYAJSOCTFBV-UHFFFAOYSA-N 4-phenyl-1,3-benzothiazole Chemical compound C1=CC=C2SC=NC2=C1C1=CC=CC=C1 RILRYAJSOCTFBV-UHFFFAOYSA-N 0.000 description 1
- NTFMLYSGIKHECT-UHFFFAOYSA-N 4-phenyl-1,3-oxazole Chemical compound O1C=NC(C=2C=CC=CC=2)=C1 NTFMLYSGIKHECT-UHFFFAOYSA-N 0.000 description 1
- MLBGDGWUZBTFHT-UHFFFAOYSA-N 4-phenyl-1,3-selenazole Chemical compound [se]1C=NC(C=2C=CC=CC=2)=C1 MLBGDGWUZBTFHT-UHFFFAOYSA-N 0.000 description 1
- KXCQDIWJQBSUJF-UHFFFAOYSA-N 4-phenyl-1,3-thiazole Chemical compound S1C=NC(C=2C=CC=CC=2)=C1 KXCQDIWJQBSUJF-UHFFFAOYSA-N 0.000 description 1
- QMUXKZBRYRPIPQ-UHFFFAOYSA-N 5,6-dimethyl-1,3-benzothiazole Chemical compound C1=C(C)C(C)=CC2=C1SC=N2 QMUXKZBRYRPIPQ-UHFFFAOYSA-N 0.000 description 1
- RWNMLYACWNIEIG-UHFFFAOYSA-N 5,6-dimethyl-1,3-benzoxazole Chemical compound C1=C(C)C(C)=CC2=C1OC=N2 RWNMLYACWNIEIG-UHFFFAOYSA-N 0.000 description 1
- QDJLLCBDLMEGEI-UHFFFAOYSA-N 5-(2-phenylethyl)-1,3-benzothiazole Chemical compound C=1C=C2SC=NC2=CC=1CCC1=CC=CC=C1 QDJLLCBDLMEGEI-UHFFFAOYSA-N 0.000 description 1
- ODSGKKPRKQLYDA-UHFFFAOYSA-N 5-(trifluoromethyl)-1,3-benzothiazole Chemical compound FC(F)(F)C1=CC=C2SC=NC2=C1 ODSGKKPRKQLYDA-UHFFFAOYSA-N 0.000 description 1
- KFDDRUWQFQJGNL-UHFFFAOYSA-N 5-bromo-1,3-benzothiazole Chemical compound BrC1=CC=C2SC=NC2=C1 KFDDRUWQFQJGNL-UHFFFAOYSA-N 0.000 description 1
- PGOGTWDYLFKOHI-UHFFFAOYSA-N 5-bromo-1,3-benzoxazole Chemical compound BrC1=CC=C2OC=NC2=C1 PGOGTWDYLFKOHI-UHFFFAOYSA-N 0.000 description 1
- DUMYZVKQCMCQHJ-UHFFFAOYSA-N 5-chloro-1,3-benzoselenazole Chemical compound ClC1=CC=C2[se]C=NC2=C1 DUMYZVKQCMCQHJ-UHFFFAOYSA-N 0.000 description 1
- YTSFYTDPSSFCLU-UHFFFAOYSA-N 5-chloro-1,3-benzothiazole Chemical compound ClC1=CC=C2SC=NC2=C1 YTSFYTDPSSFCLU-UHFFFAOYSA-N 0.000 description 1
- VWMQXAYLHOSRKA-UHFFFAOYSA-N 5-chloro-1,3-benzoxazole Chemical compound ClC1=CC=C2OC=NC2=C1 VWMQXAYLHOSRKA-UHFFFAOYSA-N 0.000 description 1
- NHUCWAWLNRUVMN-UHFFFAOYSA-N 5-chloro-6-nitro-1,3-benzoselenazole Chemical compound C1=C(Cl)C([N+](=O)[O-])=CC2=C1N=C[se]2 NHUCWAWLNRUVMN-UHFFFAOYSA-N 0.000 description 1
- OCARCLRLOLFHPY-UHFFFAOYSA-N 5-chloro-6-nitro-1,3-benzothiazole Chemical compound C1=C(Cl)C([N+](=O)[O-])=CC2=C1N=CS2 OCARCLRLOLFHPY-UHFFFAOYSA-N 0.000 description 1
- GWKNDCJHRNOQAR-UHFFFAOYSA-N 5-ethoxy-1,3-benzothiazole Chemical compound CCOC1=CC=C2SC=NC2=C1 GWKNDCJHRNOQAR-UHFFFAOYSA-N 0.000 description 1
- MHWNEQOZIDVGJS-UHFFFAOYSA-N 5-ethoxy-1,3-benzoxazole Chemical compound CCOC1=CC=C2OC=NC2=C1 MHWNEQOZIDVGJS-UHFFFAOYSA-N 0.000 description 1
- ANEKYSBZODRVRB-UHFFFAOYSA-N 5-fluoro-1,3-benzothiazole Chemical compound FC1=CC=C2SC=NC2=C1 ANEKYSBZODRVRB-UHFFFAOYSA-N 0.000 description 1
- ZRMPAEOUOPNNPZ-UHFFFAOYSA-N 5-fluoro-1,3-benzoxazole Chemical compound FC1=CC=C2OC=NC2=C1 ZRMPAEOUOPNNPZ-UHFFFAOYSA-N 0.000 description 1
- GLKZKYSZPVHLDK-UHFFFAOYSA-N 5-iodo-1,3-benzothiazole Chemical compound IC1=CC=C2SC=NC2=C1 GLKZKYSZPVHLDK-UHFFFAOYSA-N 0.000 description 1
- AHIHYPVDBXEDMN-UHFFFAOYSA-N 5-methoxy-1,3-benzoselenazole Chemical compound COC1=CC=C2[se]C=NC2=C1 AHIHYPVDBXEDMN-UHFFFAOYSA-N 0.000 description 1
- PNJKZDLZKILFNF-UHFFFAOYSA-N 5-methoxy-1,3-benzothiazole Chemical compound COC1=CC=C2SC=NC2=C1 PNJKZDLZKILFNF-UHFFFAOYSA-N 0.000 description 1
- IQQKXTVYGHYXFX-UHFFFAOYSA-N 5-methoxy-1,3-benzoxazole Chemical compound COC1=CC=C2OC=NC2=C1 IQQKXTVYGHYXFX-UHFFFAOYSA-N 0.000 description 1
- PQPFOZJCILBANS-UHFFFAOYSA-N 5-methoxybenzo[e][1,3]benzothiazole Chemical compound C12=CC=CC=C2C(OC)=CC2=C1N=CS2 PQPFOZJCILBANS-UHFFFAOYSA-N 0.000 description 1
- TTWTXOMTJQBYPG-UHFFFAOYSA-N 5-methoxybenzo[f][1,3]benzothiazole Chemical compound C1=C2C(OC)=CC=CC2=CC2=C1N=CS2 TTWTXOMTJQBYPG-UHFFFAOYSA-N 0.000 description 1
- SEBIXVUYSFOUEL-UHFFFAOYSA-N 5-methyl-1,3-benzothiazole Chemical compound CC1=CC=C2SC=NC2=C1 SEBIXVUYSFOUEL-UHFFFAOYSA-N 0.000 description 1
- UBIAVBGIRDRQLD-UHFFFAOYSA-N 5-methyl-1,3-benzoxazole Chemical compound CC1=CC=C2OC=NC2=C1 UBIAVBGIRDRQLD-UHFFFAOYSA-N 0.000 description 1
- ZYMHCFYHVYGFMS-UHFFFAOYSA-N 5-methyl-1,3-oxazole Chemical compound CC1=CN=CO1 ZYMHCFYHVYGFMS-UHFFFAOYSA-N 0.000 description 1
- JVVVSWNKYQPSEP-UHFFFAOYSA-N 5-nitro-1,3-benzoselenazole Chemical compound [O-][N+](=O)C1=CC=C2[se]C=NC2=C1 JVVVSWNKYQPSEP-UHFFFAOYSA-N 0.000 description 1
- AEUQLELVLDMMKB-UHFFFAOYSA-N 5-nitro-1,3-benzothiazole Chemical compound [O-][N+](=O)C1=CC=C2SC=NC2=C1 AEUQLELVLDMMKB-UHFFFAOYSA-N 0.000 description 1
- MNEOLRFGVQZMLA-UHFFFAOYSA-N 5-nitro-1,3-benzoxazole Chemical compound [O-][N+](=O)C1=CC=C2OC=NC2=C1 MNEOLRFGVQZMLA-UHFFFAOYSA-N 0.000 description 1
- ZOJQAWKPOGMOHO-UHFFFAOYSA-N 5-nitrobenzo[g][1,3]benzoxazole Chemical compound C12=CC=CC=C2C([N+](=O)[O-])=CC2=C1OC=N2 ZOJQAWKPOGMOHO-UHFFFAOYSA-N 0.000 description 1
- NIFNXGHHDAXUGO-UHFFFAOYSA-N 5-phenyl-1,3-benzoxazole Chemical compound C=1C=C2OC=NC2=CC=1C1=CC=CC=C1 NIFNXGHHDAXUGO-UHFFFAOYSA-N 0.000 description 1
- YJOUISWKEOXIMC-UHFFFAOYSA-N 6-bromo-1,3-benzothiazole Chemical compound BrC1=CC=C2N=CSC2=C1 YJOUISWKEOXIMC-UHFFFAOYSA-N 0.000 description 1
- AIBQGOMAISTKSR-UHFFFAOYSA-N 6-chloro-1,3-benzothiazole Chemical compound ClC1=CC=C2N=CSC2=C1 AIBQGOMAISTKSR-UHFFFAOYSA-N 0.000 description 1
- JJOOKXUUVWIARB-UHFFFAOYSA-N 6-chloro-1,3-benzoxazole Chemical compound ClC1=CC=C2N=COC2=C1 JJOOKXUUVWIARB-UHFFFAOYSA-N 0.000 description 1
- AHOIGFLSEXUWNV-UHFFFAOYSA-N 6-methoxy-1,3-benzothiazole Chemical compound COC1=CC=C2N=CSC2=C1 AHOIGFLSEXUWNV-UHFFFAOYSA-N 0.000 description 1
- FKYKJYSYSGEDCG-UHFFFAOYSA-N 6-methoxy-1,3-benzoxazole Chemical compound COC1=CC=C2N=COC2=C1 FKYKJYSYSGEDCG-UHFFFAOYSA-N 0.000 description 1
- XCJCAMHJUCETPI-UHFFFAOYSA-N 6-methyl-1,3-benzothiazol-5-ol Chemical compound C1=C(O)C(C)=CC2=C1N=CS2 XCJCAMHJUCETPI-UHFFFAOYSA-N 0.000 description 1
- IVKILQAPNDCUNJ-UHFFFAOYSA-N 6-methyl-1,3-benzothiazole Chemical compound CC1=CC=C2N=CSC2=C1 IVKILQAPNDCUNJ-UHFFFAOYSA-N 0.000 description 1
- SZWNDAUMBWLYOQ-UHFFFAOYSA-N 6-methylbenzoxazole Chemical compound CC1=CC=C2N=COC2=C1 SZWNDAUMBWLYOQ-UHFFFAOYSA-N 0.000 description 1
- KDFHXKQVTNRYLQ-UHFFFAOYSA-N 6-nitro-1,3-benzoselenazole Chemical compound [O-][N+](=O)C1=CC=C2N=C[se]C2=C1 KDFHXKQVTNRYLQ-UHFFFAOYSA-N 0.000 description 1
- QLUFBCVWKTWKBF-UHFFFAOYSA-N 6-nitro-1,3-benzothiazole Chemical compound [O-][N+](=O)C1=CC=C2N=CSC2=C1 QLUFBCVWKTWKBF-UHFFFAOYSA-N 0.000 description 1
- NNESGHWUVLNAML-UHFFFAOYSA-N 6-nitro-1,3-benzoxazole Chemical compound [O-][N+](=O)C1=CC=C2N=COC2=C1 NNESGHWUVLNAML-UHFFFAOYSA-N 0.000 description 1
- RXEDQOMFMWCKFW-UHFFFAOYSA-N 7-chloro-1,3-benzothiazole Chemical compound ClC1=CC=CC2=C1SC=N2 RXEDQOMFMWCKFW-UHFFFAOYSA-N 0.000 description 1
- PLUBSUXEOQBUFZ-UHFFFAOYSA-N 7-ethoxybenzo[g][1,3]benzothiazole Chemical compound C1=CC2=CC(OCC)=CC=C2C2=C1N=CS2 PLUBSUXEOQBUFZ-UHFFFAOYSA-N 0.000 description 1
- YVKTXAJDKKMNFM-UHFFFAOYSA-N 8-methoxybenzo[g][1,3]benzothiazole Chemical compound C12=CC(OC)=CC=C2C=CC2=C1SC=N2 YVKTXAJDKKMNFM-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical group [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 241000192041 Micrococcus Species 0.000 description 1
- ZCQWOFVYLHDMMC-UHFFFAOYSA-N Oxazole Chemical compound C1=COC=N1 ZCQWOFVYLHDMMC-UHFFFAOYSA-N 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- SJOOOZPMQAWAOP-UHFFFAOYSA-N [Ag].BrCl Chemical compound [Ag].BrCl SJOOOZPMQAWAOP-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 125000006615 aromatic heterocyclic group Chemical group 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000001420 bacteriolytic effect Effects 0.000 description 1
- AMTXUWGBSGZXCJ-UHFFFAOYSA-N benzo[e][1,3]benzoselenazole Chemical compound C1=CC=C2C(N=C[se]3)=C3C=CC2=C1 AMTXUWGBSGZXCJ-UHFFFAOYSA-N 0.000 description 1
- KXNQKOAQSGJCQU-UHFFFAOYSA-N benzo[e][1,3]benzothiazole Chemical compound C1=CC=C2C(N=CS3)=C3C=CC2=C1 KXNQKOAQSGJCQU-UHFFFAOYSA-N 0.000 description 1
- WMUIZUWOEIQJEH-UHFFFAOYSA-N benzo[e][1,3]benzoxazole Chemical compound C1=CC=C2C(N=CO3)=C3C=CC2=C1 WMUIZUWOEIQJEH-UHFFFAOYSA-N 0.000 description 1
- HJLDPBXWNCCXGM-UHFFFAOYSA-N benzo[f][1,3]benzothiazole Chemical compound C1=CC=C2C=C(SC=N3)C3=CC2=C1 HJLDPBXWNCCXGM-UHFFFAOYSA-N 0.000 description 1
- GYTPOXPRHJKGHD-UHFFFAOYSA-N benzo[f][1,3]benzoxazole Chemical compound C1=CC=C2C=C(OC=N3)C3=CC2=C1 GYTPOXPRHJKGHD-UHFFFAOYSA-N 0.000 description 1
- IEICFDLIJMHYQB-UHFFFAOYSA-N benzo[g][1,3]benzoselenazole Chemical compound C1=CC=CC2=C([se]C=N3)C3=CC=C21 IEICFDLIJMHYQB-UHFFFAOYSA-N 0.000 description 1
- IIUUNAJWKSTFPF-UHFFFAOYSA-N benzo[g][1,3]benzothiazole Chemical compound C1=CC=CC2=C(SC=N3)C3=CC=C21 IIUUNAJWKSTFPF-UHFFFAOYSA-N 0.000 description 1
- BVVBQOJNXLFIIG-UHFFFAOYSA-N benzo[g][1,3]benzoxazole Chemical compound C1=CC=CC2=C(OC=N3)C3=CC=C21 BVVBQOJNXLFIIG-UHFFFAOYSA-N 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- ZSBYCGYHRQGYNA-UHFFFAOYSA-N ethyl 1,3-benzothiazole-5-carboxylate Chemical compound CCOC(=O)C1=CC=C2SC=NC2=C1 ZSBYCGYHRQGYNA-UHFFFAOYSA-N 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- OAKJQQAXSVQMHS-UHFFFAOYSA-N hydrazine Substances NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 150000002460 imidazoles Chemical class 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- NMHMNPHRMNGLLB-UHFFFAOYSA-N phloretic acid Chemical compound OC(=O)CCC1=CC=C(O)C=C1 NMHMNPHRMNGLLB-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 125000000547 substituted alkyl group Chemical group 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 125000000020 sulfo group Chemical group O=S(=O)([*])O[H] 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Classifications
-
- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/583—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with non-fluorescent dye label
Description
本発明は抗体又は抗原を写真化学的に、且つ、
定性又は定量的に検出する免疫検査方法に関す
る。
抗原又は抗体を定性又は定量的に検出するため
に抗原又は抗体を放射性同位元素を以つて標識す
る方法や酵素を以つて標識する方法は既に用いら
れている。
また特開昭55−116259号では、本出願人によつ
て放射性同意元素や酵素の代りに分光増感色素に
より、抗原又は抗体を標識して免疫反応させ、標
識された抗原又は抗体と標識された抗原−抗体反
応物とを分離し、標識された抗原又は抗体或いは
標識された抗原−抗体反応物のどちらか一方をハ
ロゲン化銀と接触させ、分光増感色素の吸収する
波長の光で露光し、次いで、露光されたハロゲン
化銀を現像し、得られる濃度を測定することを特
徴とする微量成分の写真化学的な検査方法が提案
されており、その方法では標識物質として、写真
用分光増感色素が使用されている。
本発明も同様に標識物質として、分光増感剤を
用いるものであるが、その際複素環、即ち、酸性
核又は塩基性核にカルボキシル基を有するメロシ
アニン色素を分光増感剤として使用することを特
徴とするものである。
本発明は更に、標識物質として複素環にカルボ
キシル含有置換基を有するメロシアニン色素を使
用して、抗原又は抗体を標識し、こうして得られ
た標識抗原又は抗体を免疫反応に附し、反応生成
物と未反応物を分離したのち又は、分離すると同
時にそれらの何れか一方をハロゲン化銀と接触さ
せ、対応するメロシアニン色素の吸収する波長の
光で露光し、次いで、露光されたハロゲン化銀を
現像し、得られた濃度を測定することを特徴とす
る抗体又は抗原に標識物質を標識することにより
抗原又は抗体を写真化学的に検出する方法に関す
る。
本発明で使用されるメロシアニンは、次記一般
式()で示すことが出来る。
式中、pは2又は3の数値、nは1又は2の数
値を表わし、Zは5員又は6員の含窒素複素環
(塩基性核)を完成するに必要な非金属原子群、
Qも5又は6員含窒素複素環(酸性核)を形成す
るのに必要な非金属原子群を表わし、複素原子と
しては窒素、硫黄、セレンおよび酸素がある。R
はC数1〜18、好ましくは1〜7のアルキル基を
表わし、芳香族基、OH基、スルホ基等で置換さ
れていてもよい。そうしてこの一般式()に
は、カルボキシ基を含有する基が含窒素複素環に
結合している。酸性核及び塩基性核に関しては
“The Theory of the Photographic Process
(4th ed.)”Edited by T.H.James(1977、
Macmillan社刊)第8章参照。
Zを含有する塩基性核としては、チアゾール、
4−メチルチアゾール、4−フエニルチアゾー
ル、4・5−ジメチルチアゾール、4・5−ジフ
エニルチアゾール、ベンゾチアゾール4−クロル
ベンゾチアゾール、5−クロルベンゾチアゾー
ル、6−クロルベンゾチアゾール、7−クロルベ
ンゾチアゾール、5−ニトロベンゾチアゾール、
6−ニトロベンゾチアゾール、4−メチルベンゾ
チアゾール、5−メチルベンゾチアゾール、6−
メチルベンゾチアゾール、5−ブロモベンゾチア
ゾール、6−ブロモベンゾチアゾール、5−ヨー
ドベンゾチアゾール、5−フエニルベンゾチアゾ
ール、5−メトキシベンゾチアゾール、6−メト
キシベンゾチアゾール、5−エトキシベンゾチア
ゾール、5−エトキシカルボニルベンゾチアゾー
ル、5−フエネチルベンゾチアゾール、5−フル
オロベンゾチアゾール、5−クロル−6−ニトロ
ベンゾチアゾール、5−トリフルオロメチルベン
ゾチアゾール、5・6−ジメチルベンゾチアゾー
ル、5−ヒドロキシ−6−メチルベンゾチアゾー
ル、テトラヒドロベンゾチアゾール、4−フエニ
ルベンゾチアゾール、5−フエニルベンゾチアゾ
ール、ナフト[2・1−d]チアゾール、ナフト
[1・2−d]チアゾール、ナフト[2・3−
d]チアゾール、5−メトキシナフト[1・2−
d]チアゾール、7−エトキシナフト[2・1−
d]チアゾール、8−メトキシナフト[2・1−
d]チアゾール、5−メトキシナフト[2・3−
d]チアゾール、オキサゾール、4−メチルオキ
サゾール、4−ニトロオキサゾール、5−メチル
オキサゾール、4−フエニルオキサゾール、4・
5−ジフエニルオキサゾール、4−エチルオキサ
ゾール、ベンズオキサゾール、5−クロルベンズ
オキサゾール、5−メチルベンズオキサゾール、
5−ブロムベンズオキサゾール、5−フルオロベ
ンズオキサゾール、5−フエニルベンズオキサゾ
ール、5−メトキシベンズオキサゾール、5−ニ
トロベンズオキサゾール、5−トリフルオロベン
ズオキサゾール、5−ヒドロキシベンズオキサゾ
ール、6−メチルベンズオキサゾール、6−クロ
ルベンズオキサゾール、6−ニトロベンズオキサ
ゾール、6−メトキシベンズオキサゾール、6−
ヒドロキシベンズオキサゾール、5・6−ジメチ
ルベンズオキサゾール、4・6−ジメチルベンズ
オキサゾール、5−エトキシベンズオキサゾー
ル、ナフト[2・1−d]オキサゾール、ナフト
[1・2−d]オキサゾール、ナフト[2・3−
d]オキサゾール、5−ニトロナフト[2・1−
d]オキサゾール、4−メチルセレナゾール、4
−ニトロセレナゾール、4−フエニルセレナゾー
ル、ベンゾセレナゾール、5−クロルベンゾセレ
ナゾール、5−ニトロベンゾセレナゾール、5−
メトキシベンゾセレナゾール、5−ヒドロキシベ
ンゾセレナゾール、6−ニトロベンゾセレナゾー
ル、5−クロル−6−ニトロベンゾセレナゾー
ル、ナフト[2・1−d]セレナゾール、ナフト
[1・2−d]セレナゾール、1−アルキルイミ
ダゾール、1−アルキル−4−フエニルイミダゾ
ール、1−アルキルベンズイミダゾール、1−ア
ルキル−5−クロルベンズイミダゾール、1−ア
ルキル−5・6−ジクロル−ベンズイミダゾー
ル、1−アルキル−5−メトキシベンズイミダゾ
ール、1−アルキル−5−シアノベンズイミダゾ
ール、1−アルキル−5−フルオロベンズイミダ
ゾール、1−アルキル−5−トリフルオロメチル
ベンズイミダゾール、1−アルキルナフト[1・
2−d]イミダゾール、1−アリール−5・6−
ジクロルベンズイミダゾール、1−アリール−5
−クロルベンズイミダゾール、1−アリールイミ
ダゾール、1−アリールベンズイミダゾール、1
−アリール−5−クロルベンズイミダゾール、1
−アリール−5・6−ジクロルベンズイミダゾー
ル、1−アリール−5−メトキシベンズイミダゾ
ール、1−アリール−5−シアノベンズイミダゾ
ール、1−アリールナフト[1・2−d]イミダ
ゾール等が望ましく、その際、アルキル基は、メ
チル、エチル、プロピル、イソプロピル、ブチル
の外、2−ヒドロキシアルキル、3−ヒドロキシ
プロピル等であり、アリールは、フエニル、ハロ
ゲン、置換フエニル、メチル置換フエニル、メト
キシ置換フエニル基を表わす。塩基性核として
は、更に、2−ピリジン、4−ピリジン、5−メ
チル−2−ピリジン、3−メチル−4−ピリジン
など、キノリン核(例えば、2−キノリン、3−
メチル−2−キノリン、5−エチル−2−キノリ
ン、6−メチル−2−キノリン、6−ニトロ−2
−キノリン、8−フルオロ−2−キノリン、6−
メトキシ−2−キノリン、6−ヒドロキシ−2−
キノリン、8−クロロ−2−キノリン、4−キノ
リン、6−エトキシ−4−キノリン、6−ニトロ
−4−キノリン、8−クロロ−4−キノリン、8
−フルオロ−4−キノリン、8−メチル−4−キ
ノリン、8−メトキシ−4−キノリン)がある。
Rで示される残基としては、メチル、エチル、
プロピル、ブチル、ベンジル、β−フエニルエチ
ル、2−ヒドロキシエチル、2−メトキシエチ
ル、2−(2−メトキシエトキシ)エチル、カル
ボキシメチル、2−カルボキシエチル、3−カル
ボキシプロピル、4−カルボキシブチル、2−ス
ルホエチル、3−スルホプロピル、3−スルホブ
チル、4−スルホブチル、2−(ピロリジン−2
−オン−1−イル)エチル、テトラヒドロフルフ
リル、2−アセトキシエチル、カルボメトキシメ
チル、2−メタンスルホニルアミノエチルなどが
ある。
一般式()に含有されるべきカルボキシ基を
含有する基の代表例としては、−COOH、−
CH2COOH、−(CH2)2COOH、−NH−
CH2COOH、−NH−CH2CH2COOH、
The present invention provides photochemical photochemical analysis of antibodies or antigens, and
This invention relates to an immunoassay method for qualitative or quantitative detection. In order to qualitatively or quantitatively detect antigens or antibodies, methods of labeling antigens or antibodies with radioisotopes and enzymes have already been used. Furthermore, in JP-A No. 55-116259, the present applicant labeled an antigen or antibody with a spectral sensitizing dye instead of a radioactive element or an enzyme and caused an immunoreaction, and the labeled antigen or antibody was labeled with a spectral sensitizing dye. The labeled antigen-antibody reaction product is separated, and either the labeled antigen or antibody or the labeled antigen-antibody reaction product is brought into contact with silver halide and exposed to light at a wavelength absorbed by the spectral sensitizing dye. A photochemical inspection method for trace components has been proposed, which is characterized by developing the exposed silver halide and measuring the resulting density. Sensitizing dyes are used. The present invention similarly uses a spectral sensitizer as a labeling substance, but in this case, it is preferable to use a merocyanine dye having a carboxyl group in a heterocyclic ring, that is, an acidic nucleus or a basic nucleus, as a spectral sensitizer. This is a characteristic feature. The present invention further includes labeling an antigen or antibody using a merocyanine dye having a carboxyl-containing substituent on the heterocycle as a labeling substance, and subjecting the thus obtained labeled antigen or antibody to an immunoreaction to form a reaction product. After separating the unreacted substances, or simultaneously with the separation, either one of them is brought into contact with silver halide, exposed to light of a wavelength that is absorbed by the corresponding merocyanine dye, and then the exposed silver halide is developed. , relates to a method for photochemically detecting an antigen or antibody by labeling the antibody or antigen with a labeling substance, the method comprising measuring the concentration obtained. The merocyanine used in the present invention can be represented by the following general formula (). In the formula, p represents a numerical value of 2 or 3, n represents a numerical value of 1 or 2, and Z is a group of nonmetallic atoms necessary to complete a 5- or 6-membered nitrogen-containing heterocycle (basic nucleus);
Q also represents a nonmetallic atomic group necessary to form a 5- or 6-membered nitrogen-containing heterocycle (acidic nucleus), and the heteroatoms include nitrogen, sulfur, selenium, and oxygen. R
represents an alkyl group having 1 to 18 carbon atoms, preferably 1 to 7 carbon atoms, and may be substituted with an aromatic group, an OH group, a sulfo group, etc. In this general formula (), a group containing a carboxy group is bonded to a nitrogen-containing heterocycle. Regarding acidic nuclei and basic nuclei, “The Theory of the Photographic Process”
(4th ed.)” Edited by THJames (1977,
See Chapter 8 (published by Macmillan). As the basic nucleus containing Z, thiazole,
4-Methylthiazole, 4-phenylthiazole, 4,5-dimethylthiazole, 4,5-diphenylthiazole, benzothiazole 4-chlorobenzothiazole, 5-chlorobenzothiazole, 6-chlorobenzothiazole, 7-chlorobenzo Thiazole, 5-nitrobenzothiazole,
6-nitrobenzothiazole, 4-methylbenzothiazole, 5-methylbenzothiazole, 6-
Methylbenzothiazole, 5-bromobenzothiazole, 6-bromobenzothiazole, 5-iodobenzothiazole, 5-phenylbenzothiazole, 5-methoxybenzothiazole, 6-methoxybenzothiazole, 5-ethoxybenzothiazole, 5-ethoxy Carbonylbenzothiazole, 5-phenethylbenzothiazole, 5-fluorobenzothiazole, 5-chloro-6-nitrobenzothiazole, 5-trifluoromethylbenzothiazole, 5,6-dimethylbenzothiazole, 5-hydroxy-6- Methylbenzothiazole, tetrahydrobenzothiazole, 4-phenylbenzothiazole, 5-phenylbenzothiazole, naphtho[2,1-d]thiazole, naphtho[1,2-d]thiazole, naphtho[2,3-
d] Thiazole, 5-methoxynaphtho[1,2-
d] Thiazole, 7-ethoxynaphtho[2.1-
d] Thiazole, 8-methoxynaphtho[2.1-
d] Thiazole, 5-methoxynaphtho[2,3-
d] Thiazole, oxazole, 4-methyloxazole, 4-nitrooxazole, 5-methyloxazole, 4-phenyloxazole, 4.
5-diphenyloxazole, 4-ethyloxazole, benzoxazole, 5-chlorobenzoxazole, 5-methylbenzoxazole,
5-bromobenzoxazole, 5-fluorobenzoxazole, 5-phenylbenzoxazole, 5-methoxybenzoxazole, 5-nitrobenzoxazole, 5-trifluorobenzoxazole, 5-hydroxybenzoxazole, 6-methylbenzoxazole, 6-chlorobenzoxazole, 6-nitrobenzoxazole, 6-methoxybenzoxazole, 6-
Hydroxybenzoxazole, 5,6-dimethylbenzoxazole, 4,6-dimethylbenzoxazole, 5-ethoxybenzoxazole, naphtho[2,1-d]oxazole, naphtho[1,2-d]oxazole, naphtho[2, 3-
d] Oxazole, 5-nitronaphtho[2.1-
d] Oxazole, 4-methylselenazole, 4
-Nitroselenazole, 4-phenylselenazole, benzoselenazole, 5-chlorobenzoselenazole, 5-nitrobenzoselenazole, 5-
Methoxybenzoselenazole, 5-hydroxybenzoselenazole, 6-nitrobenzoselenazole, 5-chloro-6-nitrobenzoselenazole, naphtho[2,1-d]selenazole, naphtho[1,2-d]selenazole, 1-alkylimidazole, 1-alkyl-4-phenylimidazole, 1-alkylbenzimidazole, 1-alkyl-5-chlorobenzimidazole, 1-alkyl-5,6-dichlorobenzimidazole, 1-alkyl-5- Methoxybenzimidazole, 1-alkyl-5-cyanobenzimidazole, 1-alkyl-5-fluorobenzimidazole, 1-alkyl-5-trifluoromethylbenzimidazole, 1-alkylnaphtho[1.
2-d] imidazole, 1-aryl-5,6-
Dichlorobenzimidazole, 1-aryl-5
-Chlorbenzimidazole, 1-arylimidazole, 1-arylbenzimidazole, 1
-aryl-5-chlorobenzimidazole, 1
-Aryl-5,6-dichlorobenzimidazole, 1-aryl-5-methoxybenzimidazole, 1-aryl-5-cyanobenzimidazole, 1-arylnaphtho[1,2-d]imidazole, etc. are preferred; , alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, 2-hydroxyalkyl, 3-hydroxypropyl, etc., and aryl represents phenyl, halogen, substituted phenyl, methyl substituted phenyl, methoxy substituted phenyl group. . Basic nuclei include 2-pyridine, 4-pyridine, 5-methyl-2-pyridine, 3-methyl-4-pyridine, and quinoline nuclei (e.g., 2-quinoline, 3-pyridine).
Methyl-2-quinoline, 5-ethyl-2-quinoline, 6-methyl-2-quinoline, 6-nitro-2
-quinoline, 8-fluoro-2-quinoline, 6-
Methoxy-2-quinoline, 6-hydroxy-2-
Quinoline, 8-chloro-2-quinoline, 4-quinoline, 6-ethoxy-4-quinoline, 6-nitro-4-quinoline, 8-chloro-4-quinoline, 8
-fluoro-4-quinoline, 8-methyl-4-quinoline, 8-methoxy-4-quinoline). The residues represented by R include methyl, ethyl,
Propyl, butyl, benzyl, β-phenylethyl, 2-hydroxyethyl, 2-methoxyethyl, 2-(2-methoxyethoxy)ethyl, carboxymethyl, 2-carboxyethyl, 3-carboxypropyl, 4-carboxybutyl, 2- Sulfoethyl, 3-sulfopropyl, 3-sulfobutyl, 4-sulfobutyl, 2-(pyrrolidine-2
-on-1-yl)ethyl, tetrahydrofurfuryl, 2-acetoxyethyl, carbomethoxymethyl, 2-methanesulfonylaminoethyl, and the like. Representative examples of groups containing a carboxy group to be included in general formula () include -COOH, -
CH 2 COOH, −(CH 2 ) 2 COOH, −NH−
CH2COOH , -NH- CH2CH2COOH ,
【式】【formula】
【式】−
NHCOCH2CH2COOH、
−NHCOCH2CH2CH2COOH、
[Formula] −NHCOCH 2 CH 2 COOH, −NHCOCH 2 CH 2 CH 2 COOH,
【式】【formula】
【式】【formula】
【式】 −CONHCH2COOH、[Formula] −CONHCH 2 COOH,
【式】− C6H4−COOH[Formula] −C 6 H 4 −COOH
【式】【formula】
【式】
−CONHCH2CH2COOH、−CO
(NHCH2CO)2OH、−CO(NHCH2CO)3OH
−C6H4−CH2COOH、−C6H4CH2CH2COOH、
[Formula] −CONHCH 2 CH 2 COOH, −CO
(NHCH 2 CO) 2 OH, −CO (NHCH 2 CO) 3 OH −C 6 H 4 −CH 2 COOH, −C 6 H 4 CH 2 CH 2 COOH,
で示される化合物を安定化剤として混在せしめる
ときは分光増感色素により、標識せられた抗原又
は抗体は、水溶液中でも安定性が高く、本発明の
実施に際し好都合である。
また、本発明の写真化学的な検査法を実施する
ため、ハロゲン化銀を現像する際一般式(H)
式中R1は、アリール基又は置換アリール基R2
は水素、アルキル基、置換アルキル基、アリール
基又は置換アリール基で示されるヒドラジン化合
物を共存せしめるときは、現象銀又は発色色素の
光学濃度が増大し、測定に際し好都合である。そ
の際、濃度増強剤は、現像操作以前の何れの工程
で添加されてもよい。
以下、参考例により、N・N′−ジシクロヘキ
シルカルボジイミドを用いたN−ヒドロキシコハ
ク酸イミドエステルの合成について、本発明の分
光増感色素の反応性を示す。
参考例
色素(−1)によるフエニルアラニンの修飾
色素(−1)3.3g(10mmol)とN−ヒドロ
キシコハク酸イミド1.15g(10mmol)を60mlの
DMFに溶解し、5℃に冷却した。
N・N′−ジクロヘキシルカルボジイミド2.5g
(12.1mmol)を加え、5時間撹拌した。更に、一
夜冷蔵庫中に放置した。
析出したN・N′−ジシクロヘキシル尿素を
取し、100mlのDMFで洗浄した。液および洗液
を合わせ、これに水30mlを加えて析出した結晶を
取した。ここで得られたN−ヒドロキシコハク
酸アミドエステルは、2.5g(収率58.5%)であ
つた。
この活性エステル213.5mg(500μmol)を15ml
のDMFに溶解し、これをフエニルアラニン200mg
(1.2mmol)と0.02Mトリス塩酸緩衝液(PH7.5)
10mlとからなる溶液中に室温下約1時間で滴下し
た。その後、2時間撹拌し、一夜放置した後、、
反応混合物を水100ml中に注ぎ、析出した結晶を
取した。水洗後、メタノール/塩化メチレン=
1/1で再結晶し、108mg(収率42.8%)の目的
化合物を得た。
マススペクトル(FD)m/z=477(M+)同様
にして、化合物(−2)を使用して、フエニル
アラニンを修飾して、好収率で対応する修飾化合
物をえることができた。
実施例 1
精製ブタインシユリン(シグマ社製)20mg
(3.5μmole)を4M尿素1mlに溶解し、さらに
DMF(N・N−ジメチルホルムアミド)8mlを
加え、氷冷下(0〜4℃)で撹拌する(A液)。
色素(−2)1mg(2.7μmole)DMF1mlに溶
解したものを3組用意し、−15〜−20℃に冷却下
にクロロギ酸イソブチル各1μ及びトリエチル
アミン各0.5μを添加し、次に、同じく冷却下
にN−ヒドロキシサクシンイミド各0.5mgを添加
する(B液)。次に、A液に氷冷撹拌下に、上記
B液を5分間隔で添加し、氷冷下30分間、室温30
分間反応させた後に、0.2Nアンモニア水で平衡
化したセフアデツクスG−10カラムで脱塩後、凍
結乾燥して色素標識インシユリンを得た。収量
21.5mg(収率94%)、λ2%SDS nax=712nm ε
712om
≒8.9×10(約2モル色素/モルインシユリン)
本標識物のアミノ末端分析において、ブタイン
シユリンのアミノ末端であるグリシン及びフエニ
ルアラニンは検出されなかつた。また、セフアデ
ツクスG−50(1%SDSで平衡化)カラムのクロ
マトグラフイーにおいて、本標識物は単一ピーク
を示した。この色素にインシユリンは、塩臭化銀
乳剤(臭化物含量70%)に対し、約760nmを極
大として分光増感した。
実施例 2
ヒトリゾチーム(白血病患者の尿より精製)50
mgを2mlの2M尿素に溶解し、さらにDMFを6ml
加え、氷冷下(0〜4℃)で撹拌する。色素(
−2)各2mgを3本の小試験管に採取し、各2ml
のDMFを加えて溶解する。これを−15〜−20℃
に冷却下にクロロギ酸イソブチル各2μ及びト
リエチルアミン各1μを添加し、活性化する。
次に、上記ヒトリゾチーム溶液を、氷冷・撹拌し
ながら、活性化した色素(−2)を5分間隔で
添加・反応させる。氷冷下30分反応させた後、
0.2Nアンモニア水で平衡化したセフアデツクス
G−10カラムで脱塩後、凍結乾燥して色素標識ヒ
トリゾチームを得た。収量約45mg(収率88%)。
λ3%SDS nax=712nm ε712om≒4.6×104
本標識物のアミノ末端分析において、ヒトリゾ
チームのアミノ末端であるリジンは、検出されな
かつた。また、セフアデツクスG−50(1%SDS
で平衡化)カラムでのクロマトにより、本標品
は、単一ピークを与えた。また、ミクロコツカス
−リゾデイクテイカス(Micrococcus
Lysodeikticus)の菌体を基質にした溶菌活性に
おいて、未修飾の酵素とほぼ同等の活性を示し
た。
また、この色素標識ヒドリゾチームは、
AgBrCl乳剤(Br80モル%平均粒子サイズ0.8μ
m)に対しておよそ760nmを極大として、分光
増感性を示した。
実施例 3
実施例1に記載の色素(−2)により、標識
されたブタインシユリンと抗ブタインシユリンモ
ルモツト血清及び抗モルモツトIgGウサギ血清を
それぞれ第一抗体、第二抗体として用いた二抗体
法で、種々の濃度の標準インシユリンに対する検
量線を下記の方法で作成した。
種々の濃度(0.2ng〜12.8ng/ml)の標準イン
シユリン溶液0.1mlを小試験管に分注し、
0.1MNaCl1%牛血清アルブミン(BSA)含有の
0.1Mトリス塩酸緩衝液PH8.5(C液)各0.4mlを加
える。さらに、あらかじめ力価を定めた抗ブタイ
ンシユリンモルモツト血清の稀釈液各0.1mlを加
え、ついで色素(−2)標識インシユリンをC
液で溶解稀釈したものを各0.1ml加え、よく撹拌
したのち、4℃で16時間放置する。次に、抗モル
モツトIgGウサギ血清の稀釈液を0.1ml加え、充分
撹拌しさらに、4℃で24時間反応させる。生じた
沈澱物を遠心分離(3000rpm10分間)し、各上澄
を未露光のAgBrCl(Br80モル%、平均粒子サイ
ズ0.8μm)をTAC支持体に塗布したフイルム上
の5mmφの面積に20μ滴下し、10分間放置後、
市販のストロボ(ガイドNo.56)を用い、富士フ
イルム製SC−66フイルターを通して、30cmの距
離より露光(105lux×10-3秒に相当)、下記処方
の現像液Aにより、20℃10分間現像した後、常法
により定着、水洗、乾燥して得られたフイルム上
の黒化濃度を富士フイルム製写真濃度計にて測定
を行ない、以下の結果を得た(各抗血清標識イン
シユリンの稀釈は、標準インシユリン濃度12.8n
g/mlの場合の現像銀の黒化濃度が2.0〜2.5の間
になるように定めた)。
現像液A
メトール 3.1g
亜硫酸ソーダ 45g
ハイドロキノン 12g
無水炭酸ソーダー 67.5g
KBr 1.9g
水を加えて
表 1 インシユリン濃度(ng/ml)
黒化濃度
0 0.21
0.2 0.45
0.4 0.72
0.8 1.01
1.6 1.35
3.2 1.68
6.4 1.93
12.8 2.23
実施例 4
実施例1の色素(−2)の代りに、色素(
−5)を用いて、同様にして色素標識ブタインシ
ユリンを得た。このとき、収量19.7mg(収率86
%)、λ2%SDS nax=624nm、ε624=2.00×105
(約2
モル色素/モルインシユリン)であつた。
次に、得られた標識ブタインシユリンを用い
て、実施例3と同様に、二抗体法によつて種々の
濃度の標準インシユリンに対する満足な検量線を
得た。
When a compound represented by the following formula is mixed as a stabilizer, the antigen or antibody labeled with a spectral sensitizing dye has high stability even in an aqueous solution, which is advantageous in carrying out the present invention. In addition, in order to carry out the photochemical inspection method of the present invention, when developing silver halide, the general formula (H) In the formula, R 1 is an aryl group or a substituted aryl group R 2
When a hydrazine compound represented by hydrogen, an alkyl group, a substituted alkyl group, an aryl group, or a substituted aryl group is allowed to coexist, the optical density of the phenomenon silver or coloring dye increases, which is convenient for measurement. In this case, the density enhancer may be added at any step before the development operation. Hereinafter, reference examples will be used to demonstrate the reactivity of the spectral sensitizing dye of the present invention in the synthesis of N-hydroxysuccinimide ester using N·N'-dicyclohexylcarbodiimide. Reference example Modification of phenylalanine with dye (-1) Add 3.3 g (10 mmol) of dye (-1) and 1.15 g (10 mmol) of N-hydroxysuccinimide to 60 ml of
Dissolved in DMF and cooled to 5°C. N・N'-dichlorohexylcarbodiimide 2.5g
(12.1 mmol) was added and stirred for 5 hours. Furthermore, it was left in the refrigerator overnight. The precipitated N·N'-dicyclohexyl urea was collected and washed with 100 ml of DMF. The liquid and washing liquid were combined, 30 ml of water was added to the mixture, and the precipitated crystals were collected. The amount of N-hydroxysuccinic acid amide ester obtained here was 2.5 g (yield 58.5%). 15ml of this active ester 213.5mg (500μmol)
200mg of phenylalanine dissolved in DMF
(1.2 mmol) and 0.02 M Tris-HCl buffer (PH7.5)
It was added dropwise into a solution consisting of 10 ml at room temperature for about 1 hour. Then, after stirring for 2 hours and leaving it overnight,
The reaction mixture was poured into 100 ml of water, and the precipitated crystals were collected. After washing with water, methanol/methylene chloride =
Recrystallization was performed at a ratio of 1/1 to obtain 108 mg (yield 42.8%) of the target compound. Mass spectrum (FD) m/z = 477 (M + ) In the same manner, compound (-2) was used to modify phenylalanine to obtain the corresponding modified compound in good yield. . Example 1 Purified porcine insulin (manufactured by Sigma) 20mg
(3.5 μmole) in 1 ml of 4M urea, and
Add 8 ml of DMF (N·N-dimethylformamide) and stir under ice cooling (0 to 4°C) (liquid A).
Prepare three sets of 1 mg (2.7 μmole) of dye (-2) dissolved in 1 ml of DMF, add 1 μ each of isobutyl chloroformate and 0.5 μ each of triethylamine while cooling to −15 to −20°C, and then cool the same solution. Add 0.5 mg each of N-hydroxysuccinimide to the bottom (solution B). Next, the above-mentioned solution B was added to solution A at 5-minute intervals while stirring on ice, and the mixture was kept at room temperature for 30 minutes under ice-cooling.
After reacting for a minute, the mixture was desalted using a Sephadex G-10 column equilibrated with 0.2N aqueous ammonia and freeze-dried to obtain dye-labeled insulin. yield
21.5 mg (94% yield), λ 2 % SDS nax = 712 nm ε
712o m
≈8.9×10 (approximately 2 mol dye/mol insulin) In the amino terminal analysis of this labeled product, glycine and phenylalanine, which are the amino terminals of porcine insulin, were not detected. Furthermore, in chromatography on a Sephadex G-50 column (equilibrated with 1% SDS), this labeled substance showed a single peak. This dye, insulin, was spectrally sensitized to a silver chlorobromide emulsion (bromide content: 70%) with a maximum wavelength of about 760 nm. Example 2 Human lysozyme (purified from leukemia patient urine) 50
Dissolve mg in 2 ml of 2M urea and add 6 ml of DMF.
Add and stir under ice cooling (0 to 4°C). Pigment (
-2) Collect 2 mg of each into 3 small test tubes, and 2 ml of each
Add DMF and dissolve. -15 to -20℃
2μ each of isobutyl chloroformate and 1μ each of triethylamine are added to the mixture under cooling for activation.
Next, the activated dye (-2) is added and reacted with the human lysozyme solution at 5 minute intervals while being ice-cooled and stirred. After reacting for 30 minutes under ice cooling,
After desalting using a Sephadex G-10 column equilibrated with 0.2N aqueous ammonia, the product was freeze-dried to obtain dye-labeled human lysozyme. Yield: approximately 45 mg (yield 88%). λ 3 % SDS nax = 712 nm ε 712 o m≈4.6×10 4 In the amino terminal analysis of this labeled product, lysine, which is the amino terminal of human lysozyme, was not detected. In addition, Sephadex G-50 (1% SDS
When chromatographed on a column (equilibrated with ), this preparation gave a single peak. Also, Micrococcus - Rhizodeikteikas
In terms of bacteriolytic activity using the bacterial cells of Lysodeikticus as a substrate, the enzyme exhibited almost the same activity as the unmodified enzyme. In addition, this dye-labeled hydrisozyme
AgBrCl emulsion (Br80 mol% average grain size 0.8μ
It exhibited spectral sensitization with a maximum of approximately 760 nm relative to m). Example 3 A two-antibody method using porcine insulin labeled with the dye (-2) described in Example 1, anti-porcine insulin guinea pig serum, and anti-guinea pig IgG rabbit serum as the first antibody and second antibody, respectively, Calibration curves for standard insulin at various concentrations were created using the following method. Dispense 0.1 ml of standard insulin solutions of various concentrations (0.2 ng to 12.8 ng/ml) into small test tubes,
Contains 0.1M NaCl1% bovine serum albumin (BSA)
Add 0.4 ml each of 0.1M Tris-HCl buffer PH8.5 (Solution C). Furthermore, 0.1 ml of each diluted solution of anti-pig insulin guinea pig serum with predetermined titer was added, and then dye (-2) labeled insulin was added to C.
Add 0.1 ml of each solution dissolved and diluted with liquid, stir well, and leave at 4°C for 16 hours. Next, add 0.1 ml of diluted anti-guinea pig IgG rabbit serum, stir thoroughly, and react at 4°C for 24 hours. The resulting precipitate was centrifuged (3000 rpm for 10 minutes), and 20 μ of each supernatant was dropped onto a 5 mm diameter area on a film coated with unexposed AgBrCl (Br 80 mol%, average particle size 0.8 μm) on a TAC support. After leaving it for 10 minutes,
Using a commercially available strobe (Guide No. 56), exposure from a distance of 30 cm through a Fujifilm SC-66 filter (equivalent to 10 5 lux x 10 -3 seconds), and developing solution A with the following formulation at 20°C 10 After developing for 1 minute, the film was fixed using a conventional method, washed with water, and dried. The blackening density on the obtained film was measured using a Fujifilm photographic densitometer, and the following results were obtained (for each antiserum-labeled insulin). Dilution is standard insulin concentration 12.8n
The blackening density of the developed silver was determined to be between 2.0 and 2.5 (g/ml). Developer A Metol 3.1g Sodium sulfite 45g Hydroquinone 12g Anhydrous soda 67.5g KBr 1.9g Add water and Table 1 Insulin concentration (ng/ml) Blackening concentration 0 0.21 0.2 0.45 0.4 0.72 0.8 1.01 1.6 1.35 3.2 1.68 6.4 1.93 12.8 2.23 Example 4 In place of the dye (-2) in Example 1, the dye (
-5), dye-labeled porcine insulin was obtained in the same manner. At this time, the yield was 19.7 mg (yield: 86
%), λ 2 % SDS nax = 624 nm, ε 624 = 2.00×10 5
(about 2
Mol dye/mol insulin). Next, using the obtained labeled porcine insulin, satisfactory calibration curves for standard insulin at various concentrations were obtained by the two-antibody method in the same manner as in Example 3.
Claims (1)
環にカルボキシル基またはカルボキシル基を含む
置換基を有する置換基を結合して含有するメロシ
アニン色素を使用することを特徴とする 抗原または抗体に、分光増感色素をペプチド
結合により結合させて標識した抗原または抗体
を免疫反応させ、 測定さるべき抗原又は抗体の存在量に応じて
生成した遊離の標識された抗原又は抗体(F)と標
識された抗原−抗体反応(B)とを分離し、 標識された抗原又は抗体、或いは標識された
抗原−抗体反応物のどちらか一方をハロゲン化
銀と接触させ、 分光増感色素の吸収する波長の光で露光し、 露光されたハロゲン化銀を現像し、 得られる現像された銀又は発色色素の量、或
いは現像された銀又は発色色素の光学濃度を測
定することによる 抗原または抗体の写真化学的免疫検査方法。[Scope of Claims] 1. An antigen characterized in that a merocyanine dye containing a carboxyl group or a substituent having a carboxyl group-containing substituent bonded to a heterocyclic ring is used as a spectral sensitizing dye as a labeling substance. Alternatively, a spectral sensitizing dye is bound to the antibody via a peptide bond, and the labeled antigen or antibody is subjected to immunoreaction, and free labeled antigen or antibody (F) is generated depending on the amount of the antigen or antibody to be measured. The labeled antigen-antibody reaction (B) is separated, and either the labeled antigen or antibody, or the labeled antigen-antibody reaction product is brought into contact with silver halide to absorb the spectral sensitizing dye. the amount of developed silver or color dye obtained, or the optical density of the developed silver or color dye. Photo chemical immunoassay method.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP12059580A JPS5745456A (en) | 1980-09-02 | 1980-09-02 | Method for photographic-chemical immunoassay |
DE8181106822T DE3170134D1 (en) | 1980-09-02 | 1981-09-01 | Method for immunochemical measurement |
EP81106822A EP0047470B1 (en) | 1980-09-02 | 1981-09-01 | Method for immunochemical measurement |
US06/506,225 US4473652A (en) | 1980-09-02 | 1983-06-22 | Method and immunochemical measurement |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP12059580A JPS5745456A (en) | 1980-09-02 | 1980-09-02 | Method for photographic-chemical immunoassay |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5745456A JPS5745456A (en) | 1982-03-15 |
JPS6223825B2 true JPS6223825B2 (en) | 1987-05-25 |
Family
ID=14790144
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP12059580A Granted JPS5745456A (en) | 1980-09-02 | 1980-09-02 | Method for photographic-chemical immunoassay |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5745456A (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH083127B2 (en) * | 1990-12-28 | 1996-01-17 | 株式会社神戸製鋼所 | Method for producing high strength galvanized steel sheet with excellent workability |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55116259A (en) * | 1979-03-01 | 1980-09-06 | Fuji Photo Film Co Ltd | Microimmunoassay method |
-
1980
- 1980-09-02 JP JP12059580A patent/JPS5745456A/en active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55116259A (en) * | 1979-03-01 | 1980-09-06 | Fuji Photo Film Co Ltd | Microimmunoassay method |
Also Published As
Publication number | Publication date |
---|---|
JPS5745456A (en) | 1982-03-15 |
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