JP4083213B2 - 電気化学的免疫測定チップ - Google Patents
電気化学的免疫測定チップ Download PDFInfo
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- JP4083213B2 JP4083213B2 JP2007535937A JP2007535937A JP4083213B2 JP 4083213 B2 JP4083213 B2 JP 4083213B2 JP 2007535937 A JP2007535937 A JP 2007535937A JP 2007535937 A JP2007535937 A JP 2007535937A JP 4083213 B2 JP4083213 B2 JP 4083213B2
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- 238000003018 immunoassay Methods 0.000 title claims description 11
- 239000013076 target substance Substances 0.000 claims description 36
- -1 potassium ferricyanide Chemical compound 0.000 claims description 25
- 239000000758 substrate Substances 0.000 claims description 22
- 102000013460 Malate Dehydrogenase Human genes 0.000 claims description 17
- 108010026217 Malate Dehydrogenase Proteins 0.000 claims description 17
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims description 10
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 9
- 229940049920 malate Drugs 0.000 claims description 8
- WPUMTJGUQUYPIV-JIZZDEOASA-L disodium (S)-malate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](O)CC([O-])=O WPUMTJGUQUYPIV-JIZZDEOASA-L 0.000 claims description 6
- 235000019265 sodium DL-malate Nutrition 0.000 claims description 6
- 239000001394 sodium malate Substances 0.000 claims description 6
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 2
- YAGKRVSRTSUGEY-UHFFFAOYSA-N ferricyanide Chemical compound [Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] YAGKRVSRTSUGEY-UHFFFAOYSA-N 0.000 claims description 2
- 229910052700 potassium Inorganic materials 0.000 claims description 2
- 239000011591 potassium Substances 0.000 claims description 2
- 239000001415 potassium malate Substances 0.000 claims description 2
- SVICABYXKQIXBM-UHFFFAOYSA-L potassium malate Chemical compound [K+].[K+].[O-]C(=O)C(O)CC([O-])=O SVICABYXKQIXBM-UHFFFAOYSA-L 0.000 claims description 2
- 235000011033 potassium malate Nutrition 0.000 claims description 2
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 claims 4
- 238000006243 chemical reaction Methods 0.000 description 71
- 239000000243 solution Substances 0.000 description 58
- 239000003153 chemical reaction reagent Substances 0.000 description 50
- 102000004190 Enzymes Human genes 0.000 description 33
- 108090000790 Enzymes Proteins 0.000 description 33
- 230000001351 cycling effect Effects 0.000 description 25
- 238000005259 measurement Methods 0.000 description 23
- 230000000052 comparative effect Effects 0.000 description 19
- XJLXINKUBYWONI-NNYOXOHSSA-O NADP(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-O 0.000 description 16
- 239000012488 sample solution Substances 0.000 description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 13
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 13
- 238000010586 diagram Methods 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 239000011259 mixed solution Substances 0.000 description 8
- 239000000523 sample Substances 0.000 description 7
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 6
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 6
- 108091006149 Electron carriers Proteins 0.000 description 6
- ACFIXJIJDZMPPO-NNYOXOHSSA-N NADPH Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](OP(O)(O)=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-N 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 5
- 239000000276 potassium ferrocyanide Substances 0.000 description 5
- XOGGUFAVLNCTRS-UHFFFAOYSA-N tetrapotassium;iron(2+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+2].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] XOGGUFAVLNCTRS-UHFFFAOYSA-N 0.000 description 5
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 229950006238 nadide Drugs 0.000 description 4
- 229920000139 polyethylene terephthalate Polymers 0.000 description 4
- 239000005020 polyethylene terephthalate Substances 0.000 description 4
- 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 3
- 239000007853 buffer solution Substances 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- AWDBHOZBRXWRKS-UHFFFAOYSA-N tetrapotassium;iron(6+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+6].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] AWDBHOZBRXWRKS-UHFFFAOYSA-N 0.000 description 3
- 239000012472 biological sample Substances 0.000 description 2
- 238000004891 communication Methods 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000007650 screen-printing Methods 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- BOPGDPNILDQYTO-NNYOXOHSSA-L NADH(2-) Chemical group C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP([O-])(=O)OP([O-])(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-L 0.000 description 1
- 239000004642 Polyimide Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000006356 dehydrogenation reaction Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229920001721 polyimide Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000000941 radioactive substance Substances 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
Images
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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/001—Enzyme electrodes
- C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
- C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/904—Oxidoreductases (1.) acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
1Mのフェリシアン化カリウム溶液(1μL)、1000U/mLのジアホラーゼ溶液(6.7μL)、4Mのリンゴ酸ナトリウム溶液(7.8μL)、5mMのNADP溶液(1μL)、25000U/mLのリンゴ酸脱水素酵素溶液(1μL)、および1MのTris−HCl溶液(5μL、pH9)を混合することにより、混合溶液を調製した。この混合溶液(22.5μL)を、チップ300の試薬固定部位18上に配置した後、常温(25℃)で3時間、真空乾燥させた。これにより、図4に示す酵素サイクリング反応に関与する試薬群を、全て混合した状態で試薬固定部位18上に乾燥固定した。
図4に示す酵素サイクリング反応に関与する試薬群を、電子伝達体(フェリシアン化カリウムおよびNADP)と、酵素(リンゴ酸脱水素酵素およびジアホラーゼ)とを分離した状態で試薬固定部位18上に乾燥固定したチップ300を用いたこと以外は、比較例1と同様にして定電位測定を実施した。
図4に示す酵素サイクリング反応に関与する試薬群を、電子伝達体のうちフェリシアン化カリウムのみを酵素およびその基質から分離した状態で、試薬固定部位18上に乾燥固定したチップ300を用いたこと以外は、比較例2と同様にして定電位測定を実施した。
図4に示す酵素サイクリング反応に関与する試薬群を、電子伝達体のうちNADPのみを酵素およびその基質から分離した状態で、試薬固定部位18上に乾燥固定したチップ300を用いたこと以外は、比較例2と同様にして定電位測定を実施した。
Claims (3)
- 免疫測定法を用いて電気化学的に標的物質の量を測定するチップであって、
前記チップは、作用極および対極からなる電極、または作用極、対極、および参照極からなる電極を有し、
該チップ内に、NADPおよびNADPHから選ばれる少なくとも1種、リンゴ酸脱水素酵素、該リンゴ酸脱水素酵素の基質、フェリシアン化カリウムならびにジアホラーゼが固定され、
前記リンゴ酸脱水素酵素と、前記NADPおよび前記NADPHから選ばれる少なくとも1種と、前記基質とが混合された状態で同一の場所に固定され、
前記リンゴ酸脱水素酵素と、前記フェリシアン化カリウムとが離間した状態で固定されたチップ。 - 前記リンゴ酸脱水素酵素の前記基質が、リンゴ酸およびリンゴ酸塩から選ばれる少なくとも1種である、請求項1に記載のチップ。
- 前記リンゴ酸塩が、リンゴ酸ナトリウムおよびリンゴ酸カリウムから選ばれる少なくとも1種を含む、請求項2に記載のチップ。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2006192581 | 2006-07-13 | ||
JP2006192581 | 2006-07-13 | ||
PCT/JP2007/060447 WO2008007499A1 (fr) | 2006-07-13 | 2007-05-22 | Puce a dosage immunologique electrochimique |
Publications (2)
Publication Number | Publication Date |
---|---|
JP4083213B2 true JP4083213B2 (ja) | 2008-04-30 |
JPWO2008007499A1 JPWO2008007499A1 (ja) | 2009-12-10 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2007535937A Active JP4083213B2 (ja) | 2006-07-13 | 2007-05-22 | 電気化学的免疫測定チップ |
Country Status (3)
Country | Link |
---|---|
US (1) | US7585400B2 (ja) |
JP (1) | JP4083213B2 (ja) |
WO (1) | WO2008007499A1 (ja) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
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JP5500791B2 (ja) * | 2008-06-19 | 2014-05-21 | デンカ生研株式会社 | 新規検査方法及びそれに用いる検査キット |
JP2021089141A (ja) * | 2018-03-26 | 2021-06-10 | Phcホールディングス株式会社 | バイオセンサ |
NO345437B1 (en) | 2018-06-01 | 2021-02-01 | Prores E&P As | Mud loss treatment drilling tool and method |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5853863B2 (ja) * | 1978-07-14 | 1983-12-01 | 松下電器産業株式会社 | 酵素電極系 |
JPS59166084A (ja) * | 1983-03-11 | 1984-09-19 | Hitachi Chem Co Ltd | 安定な酵素試薬の製造法 |
JP2502666B2 (ja) | 1988-01-29 | 1996-05-29 | 松下電器産業株式会社 | バイオセンサ及びその製造方法 |
JPH0296649A (ja) * | 1988-10-04 | 1990-04-09 | Unitika Ltd | デヒドロゲナーゼ電極 |
JP3214188B2 (ja) * | 1993-10-08 | 2001-10-02 | 松下電器産業株式会社 | バイオセンサおよびその製造法 |
JPH08327852A (ja) | 1995-05-31 | 1996-12-13 | Sumitomo Electric Ind Ltd | 光ファイバカプラおよびその製造方法 |
KR0156176B1 (ko) * | 1995-06-01 | 1998-12-01 | 구자홍 | 전기화학식 면역 바이오센서 |
JPH09297121A (ja) | 1996-03-07 | 1997-11-18 | Matsushita Electric Ind Co Ltd | コレステロールセンサ |
JP2005046001A (ja) * | 2001-06-15 | 2005-02-24 | Matsushita Electric Ind Co Ltd | 脂質修飾酵素の製造方法およびバイオセンサ |
JP2004024254A (ja) * | 2002-04-30 | 2004-01-29 | Masao Umemoto | ミオイノシトールの簡易測定法 |
WO2005108968A1 (ja) * | 2004-05-12 | 2005-11-17 | Matsushita Electric Industrial Co., Ltd. | バイオセンサ、バイオセンサ用容器、およびバイオセンサ測定装置 |
-
2007
- 2007-05-22 JP JP2007535937A patent/JP4083213B2/ja active Active
- 2007-05-22 WO PCT/JP2007/060447 patent/WO2008007499A1/ja active Application Filing
- 2007-10-30 US US11/976,950 patent/US7585400B2/en active Active
Also Published As
Publication number | Publication date |
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US7585400B2 (en) | 2009-09-08 |
US20080308419A1 (en) | 2008-12-18 |
JPWO2008007499A1 (ja) | 2009-12-10 |
WO2008007499A1 (fr) | 2008-01-17 |
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