CN1661370A - High stable method of chemiluminescence accelerated by enzyme and substrate - Google Patents
High stable method of chemiluminescence accelerated by enzyme and substrate Download PDFInfo
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- CN1661370A CN1661370A CN 200410005756 CN200410005756A CN1661370A CN 1661370 A CN1661370 A CN 1661370A CN 200410005756 CN200410005756 CN 200410005756 CN 200410005756 A CN200410005756 A CN 200410005756A CN 1661370 A CN1661370 A CN 1661370A
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Abstract
The present invention relates to a high-stability enzymatic chemiluminescence method and its substrate. According to specific proportion said method uses clumiluminescence substrate A liquid and B liquid, after HRP labeled immunological reaction they are added and mixed uniformly, in which said liquid A contains luminol (5'-amino-2,3-dihydro-1,4-phthalizinedione) and reinforcing agent p-iodophenol, and the liquid B contains urea peroxide. Said invention has the advantaegs of high luminescence sensitivity, good stability, accurate result, convenient application, so that it has extensive application range.
Description
The present invention relates to a kind of high stability enzyme-catalyzed chemical luminescence method and substrate thereof.To iodophenol, oxygenant urea peroxide, form by A liquid that is mixed with respectively and B liquid by the luminol of special ratios, reinforcing agent for substrate.The A liquid of equivalent and B liquid contact with catalyzer (as horseradish peroxidase) label after the immune response, generate a kind of product of excited state by chemical reaction, and it produces luminescence phenomenon thereby the energy that discharges is transformed into photon in getting back to the process of ground state.Its advantage be have luminous sensitivity height, good stability, the result is accurate, easy to use and cost is low etc.Be applicable to full-automatic, semi-automatic and manual analysis.Can be widely used in clinical examination, biological study, legal medical expert identifies, in farming and animal husbandry and the environmental science to the detections of biomacromolecules such as nucleic acid and antigen-antibody.
Chemiluminescence immune analysis method becomes the outstanding person in the present external diagnosis reagent detection method because of environmentally safe, to comprehensive advantage and potentiality such as human body is nontoxic.
The objective of the invention is further to improve the stability of chemiluminescence immune assay substrate, for the suitability for industrialized production of product is brought bigger convenience and dirigibility with using.
Subject matter of the present invention:
1, substrate solution preparation: the Tris-HCl damping fluid of 0.2mol/L pH 8.6 is prepared A liquid and B liquid respectively: A liquid contains the luminol of 5mg/ml, and the reinforcing agent of 0.33mg/ml is to iodophenol; B liquid contains the urea peroxide of 0.9g/ml;
2, storage stability height.A liquid and B liquid store respectively.Preserved the term of validity 2 years for 2～8 ℃.
3, reaction stability height.The A liquid and the B liquid that add mixed in equal amounts in the sample hose after the immune response of HRP mark (hole), the same sample well luminous value of each time point determining stable (Fig. 1) in hour (not comprising at once) at once～3; At once～3 the same sample well measured object of each time point concentration value basically identical (Fig. 2) in hour (comprising at once).
Claims (3)
1, a kind of high stability enzyme-catalyzed chemical luminescence method and substrate thereof is characterized in that said high stability enzyme-catalyzed chemical luminescence method and substrate thereof may further comprise the steps:
(1) preparation of chemical luminous substrate A liquid and B liquid;
(2) storage stability of chemical luminous substrate;
(3) reaction stability of chemical luminous substrate.
2, according to claim 1 described high stability enzyme-catalyzed chemical luminescence method and substrate thereof, it is characterized in that:
(1) prepare A liquid and B liquid respectively with the Tris-HCl damping fluid: A liquid contains luminol and reinforcing agent to iodophenol; B liquid contains the oxygenant urea peroxide;
(2) A liquid and B liquid store respectively, and the former needs lucifuge;
(3) face with preceding A liquid and B liquid mixed in equal amounts, after the immune response of HRP mark, add in the sample hose (hole).
3, according to claim 1 described high stability enzyme-catalyzed chemical luminescence method and substrate thereof, it is characterized in that:
(1) storage stability, A liquid and B liquid store respectively, preserve the term of validity 2 years for 2～8 ℃.
(2) reaction stability is 3 hours, is included in the A liquid and the stability of its luminous value of B liquid and the stability of measured object concentration value that add mixed in equal amounts after the immune response of HRP mark in the sample hose (hole).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410005756 CN1661370A (en) | 2004-02-23 | 2004-02-23 | High stable method of chemiluminescence accelerated by enzyme and substrate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410005756 CN1661370A (en) | 2004-02-23 | 2004-02-23 | High stable method of chemiluminescence accelerated by enzyme and substrate |
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| CN1661370A true CN1661370A (en) | 2005-08-31 |
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| CN 200410005756 Pending CN1661370A (en) | 2004-02-23 | 2004-02-23 | High stable method of chemiluminescence accelerated by enzyme and substrate |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101571483B (en) * | 2008-04-29 | 2011-11-02 | 北京科美东雅生物技术有限公司 | Chemoluminescent substrate |
| CN102735679A (en) * | 2012-06-26 | 2012-10-17 | 博奥赛斯（天津）生物科技有限公司 | Kit for aldosterone chemiluminescence immunization quantitative determination and its preparation method |
| CN102890083A (en) * | 2012-09-28 | 2013-01-23 | 辽宁科骏生物有限公司 | Chemiluminescence substrate solution, kit containing same, and detection method applying chemiluminescence substrate solution or kit |
| CN103604918A (en) * | 2013-11-28 | 2014-02-26 | 中国科学院广州生物医药与健康研究院 | Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate |
| US10711185B2 (en) | 2018-07-13 | 2020-07-14 | Bio-Helix Co., Ltd. | Reagent and kit for performing chemiluminescent reaction |
| US11112366B2 (en) | 2019-03-28 | 2021-09-07 | National Taiwan Ocean University | Reagent and kit for enhancing chemiluminescent reaction |
-
2004
- 2004-02-23 CN CN 200410005756 patent/CN1661370A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101571483B (en) * | 2008-04-29 | 2011-11-02 | 北京科美东雅生物技术有限公司 | Chemoluminescent substrate |
| CN102735679A (en) * | 2012-06-26 | 2012-10-17 | 博奥赛斯（天津）生物科技有限公司 | Kit for aldosterone chemiluminescence immunization quantitative determination and its preparation method |
| CN102735679B (en) * | 2012-06-26 | 2015-07-15 | 博奥赛斯（天津）生物科技有限公司 | Kit for aldosterone chemiluminescence immunization quantitative determination and its preparation method |
| CN102890083A (en) * | 2012-09-28 | 2013-01-23 | 辽宁科骏生物有限公司 | Chemiluminescence substrate solution, kit containing same, and detection method applying chemiluminescence substrate solution or kit |
| CN103604918A (en) * | 2013-11-28 | 2014-02-26 | 中国科学院广州生物医药与健康研究院 | Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate |
| US10711185B2 (en) | 2018-07-13 | 2020-07-14 | Bio-Helix Co., Ltd. | Reagent and kit for performing chemiluminescent reaction |
| US11112366B2 (en) | 2019-03-28 | 2021-09-07 | National Taiwan Ocean University | Reagent and kit for enhancing chemiluminescent reaction |
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| PB01 | Publication | ||
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| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |