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Stable HRP (Horseradish Peroxidase) enzyme-catalyzed chemiluminescence substrate solution as well as preparation method and application thereof

An enzymatic chemiluminescence, substrate liquid technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, etc., can solve the problems of poor stability, low sensitivity, low luminescence intensity, etc., to improve stability Effect

Active Publication Date: 2016-07-27
BEIJING UNIDIAG TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0014] Based on the above problems and needs, the present invention provides a stable HRP enzymatic chemiluminescent substrate solution, a configuration method and its application. The chemiluminescent substrate solution provided by the present invention can solve the problem of the short and stable Poor performance, low luminous intensity, low sensitivity, high background

Method used

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  • Stable HRP (Horseradish Peroxidase) enzyme-catalyzed chemiluminescence substrate solution as well as preparation method and application thereof
  • Stable HRP (Horseradish Peroxidase) enzyme-catalyzed chemiluminescence substrate solution as well as preparation method and application thereof
  • Stable HRP (Horseradish Peroxidase) enzyme-catalyzed chemiluminescence substrate solution as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0074] The composition of the HRP enzymatic chemiluminescence substrate liquid of embodiment 1 of the present invention is:

[0075] The component of described A liquid is:

[0076] 0.8g / L luminol,

[0077] 1.2g / L p-imidazole phenol,

[0078] 0.2g / L sodium tetraphenylborate,

[0079] 15ml / L dimethylformamide (DMF),

[0080] 1g / L sulfobutyl-β-cyclodextrin,

[0081] 1g / L light stabilizer,

[0082] 0.25mol / L Tris buffer of pH 9.0;

[0083] The components of B are:

[0084] 3g / L polyvinylpyrrolidone (PVP),

[0085] 0.4g / L carbamide peroxide,

[0086] 0.2mol / L Tris buffer at pH 7.0.

[0087] The preparation of the luminescent substrate solution described in Embodiment 1 can be carried out according to the following process, including the following steps: the preparation of the A liquid: (1) prepare 1 L of 0.25 mol / L Tris buffer solution of pH 9.0 for standby (2) Measure 900ml of the above-mentioned Tris buffer, add 0.2g sodium tetraphenylborate and 1g sulfobutyl-β-cyclode...

Embodiment approach 2

[0092] The composition of the HRP enzymatic chemiluminescence substrate liquid of embodiment 2 of the present invention is:

[0093] The component of described A liquid is:

[0094] 0.6g / L Luminol (Luminol),

[0095] 1g / L p-imidazole phenol,

[0096] 0.4g / L sodium tetraphenylborate,

[0097] 30ml / L dimethylformamide (DMF),

[0098] 8g / L sulfobutyl-β-cyclodextrin,

[0099] 1.3g / L light stabilizer,

[0100] 0.25mol / L Tris buffer of pH8.5;

[0101] The components of B are:

[0102] 4g / L polyvinylpyrrolidone (PVP),

[0103] 0.3g / L carbamide peroxide,

[0104] 0.25mol / L Tris buffer at pH 7.0.

[0105] The preparation of the luminescent substrate solution described in embodiment 2 can be carried out according to the following process, including the following steps: the preparation of the A solution: (1) prepare 1 L of 0.25 mol / L Tris buffer solution of pH 8.5 for standby (2) Measure 900ml of the above-mentioned Tris buffer solution, add 0.4g sodium tetraphenylborate and 8g...

Embodiment approach 3

[0110] The composition of the HRP enzymatic chemiluminescence substrate liquid of embodiment 3 of the present invention is:

[0111] The component of described A liquid is:

[0112] 0.8g / L Luminol (Luminol),

[0113] 1g / L p-imidazole phenol,

[0114] 0.2g / L sodium tetraphenylborate,

[0115] 30ml / L dimethylformamide (DMF),

[0116]4g / L sulfobutyl-β-cyclodextrin,

[0117] 0.8g / L light stabilizer,

[0118] 0.25mol / L Tris buffer of pH 9.0;

[0119] The components of the B liquid are:

[0120] 3g / L polyvinylpyrrolidone (PVP),

[0121] 0.6g / L carbamide peroxide,

[0122] 0.25mol / L Tris buffer at pH 9.0.

[0123] The preparation of the luminescent substrate solution described in Embodiment 3 can be carried out according to the following process, including the following steps: the preparation of the A liquid: (1) prepare 1 L of 0.25 mol / L Tris buffer solution with a pH of 9.0 for use (2) Measure 900ml of the above-mentioned Tris buffer solution, add 0.2g sodium tetraphenylb...

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Abstract

The invention provides a stable HRP (Horseradish Peroxidase) enzyme-catalyzed chemiluminescence substrate solution as well as a preparation method and application thereof. Typically, the stable HRP enzyme-catalyzed chemiluminescence substrate solution is prepared from two components including a chemiluminescence substrate solution A and a chemiluminescence substrate solution B, wherein the solution A is prepared from luminol, p-imidazophenol, sodium tetraphenylborate, dimethylformamide (DMF), sulfobutyl-beta-cyclodextrin, a light stabilizer and a Tris buffering solution with a certain concentration; and the solution B is prepared from polyvinylpyrrolidone (PVP), urea peroxide and the Tris buffering solution. With the adoption of the chemiluminescence substrate solution provided by the invention, the problems of an existing chemiluminescence substrate that the platform phase is short, the stability is relatively poor, the luminous intensity is low, the sensitivity is low and the background is relatively high can be solved.

Description

technical field [0001] The invention relates to a composition used in the field of chemiluminescence immunoassay; specifically, the invention relates to a stable HRP enzymatic chemiluminescence substrate solution, a preparation method and application of the substrate solution. Background technique [0002] Chemiluminescence refers to a kind of light radiation phenomenon accompanied by the chemical reaction of substances, which can be divided into direct luminescence and indirect luminescence. Two substances undergo a chemical reaction to form a new substance, and the energy released by the reaction is absorbed by the molecules of the newly formed substance and transitions to an excited state, and light radiation is generated during the process of returning from the excited state to the ground state. The excess energy is released in the form of photons, a phenomenon known as chemiluminescence. [0003] Chemiluminescence immunoassay using the principle of chemiluminescence: (...

Claims

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Application Information

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IPC IPC(8): G01N21/76
CPCG01N21/76
Inventor 秦军谢元东
Owner BEIJING UNIDIAG TECH
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