Stable and efficient HRP enzymatic substrate solutions and preparation method and application thereof

A substrate solution and enzymatic technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor stability of chromogenic solution, insoluble TMB in water, and insufficient long-lasting color development, etc., to improve detection sensitivity, anti- High temperature capability and improved stability

Pending Publication Date: 2020-01-17
BEIJING BEIER BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, TMB is relatively difficult to dissolve in water, and its properties are unstable in the solution state, so it is difficult to store, which results in the poor stability of conventional TMB chromogenic solutions on the market, insufficient color development, and reduced color intensity after long-term storage. question

Method used

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  • Stable and efficient HRP enzymatic substrate solutions and preparation method and application thereof
  • Stable and efficient HRP enzymatic substrate solutions and preparation method and application thereof
  • Stable and efficient HRP enzymatic substrate solutions and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] A stable and efficient HRP enzymatic substrate solution, including substrate solution A and substrate solution B, wherein the substrate solution A includes the following final concentrations of components:

[0041] Anhydrous sodium acetate 30g / L, Tween 20 100mL / L, sodium citrate 15g / L, citric acid 20g / L, acetic acid 2.5mL / L, hydrogen peroxide 2mL / L, sodium peroxide 0.03g / L;

[0042] Substrate solution B includes components at the following final concentrations:

[0043] TMB 3g / L, dimethyl sulfoxide 10mL / L, hydrochloric acid 5mL / L, glycerin 100mL / L, sodium thiosulfate 20g / L.

[0044] Wherein, the preparation method of the substrate liquid A comprises the following steps: in the amount of 1 L, each component is weighed according to the above ratio, dissolved in distilled water respectively, and the volume is adjusted to 1 L with distilled water, and the preparation is completed;

[0045] The preparation method of substrate liquid B comprises the following steps: in the a...

Embodiment 2

[0047] A stable and efficient HRP enzymatic substrate solution, including substrate solution A and substrate solution B, wherein the substrate solution A includes the following final concentrations of components:

[0048] Anhydrous sodium acetate 6g / L, Tween 20 20mL / L, sodium citrate 3g / L, citric acid 20g / L, acetic acid 0.5mL / L, hydrogen peroxide 0.5mL / L, sodium peroxide 0.006g / L;

[0049] Substrate solution B includes components at the following final concentrations:

[0050] TMB 0.2g / L, dimethyl sulfoxide 2mL / L, hydrochloric acid 0.5mL / L, glycerol 10mL / L, sodium thiosulfate 2g / L.

[0051] Wherein, the preparation method of the substrate liquid A comprises the following steps: in the amount of 1 L, each component is weighed according to the above ratio, dissolved in distilled water respectively, and the volume is adjusted to 1 L with distilled water, and the preparation is completed;

[0052] The preparation method of substrate liquid B comprises the following steps: in the ...

Embodiment 3

[0054] A stable and efficient HRP enzymatic substrate solution, including substrate solution A and substrate solution B, wherein the substrate solution A includes the following final concentrations of components:

[0055] Anhydrous sodium acetate 15g / L, Tween 20 60mL / L, sodium citrate 8g / L, citric acid 10g / L, acetic acid 1.25mL / L, hydrogen peroxide 1mL / L, sodium peroxide 0.015g / L;

[0056] Substrate solution B includes components at the following final concentrations:

[0057] TMB 1g / L, dimethyl sulfoxide 5mL / L, hydrochloric acid 2mL / L, glycerin 50mL / L, sodium thiosulfate 10g / L.

[0058] Wherein, the preparation method of the substrate liquid A comprises the following steps: in the amount of 1 L, each component is weighed according to the above ratio, dissolved in distilled water respectively, and the volume is adjusted to 1 L with distilled water, and the preparation is completed;

[0059] The preparation method of substrate liquid B comprises the following steps: in the amo...

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Abstract

Embodiments of the invention relate to the technical field of in vitro clinical detection, in particular to stable and efficient HRP enzymatic substrate solutions and a preparation method and application thereof. The HRP enzymatic substrate solutions provided by the embodiments of the invention comprise a substrate solution A composed of sodium acetate anhydrous of 6-30g / L, Tween 20 of 20-100mL / L,sodium citrate of 3-15g / L, citric acid of 4-20g / L, acetic acid of 0.5-2.5mL / L, hydrogen peroxide of 0.5-2mL / L, and sodium peroxide of 0.006-0.03g / L; and a substrate solution B composed of TMB of 0.2-3g / L, dimethyl sulfoxide of 2-10mL / L, hydrochloric acid of 0.5-5mL / L, glycerin of 10-100mL / L and sodium thiosulfate 2-20g / L. The HRP enzymatic substrate solutions provided in the embodiments of the invention have high color developing strength and high detection sensitivity without increasing the background, have the color rendering effect lasting 1 hour, can be placed stably for 3 years at 2-8 DEG C, has strong high temperature resistance capability, can resist the influence of high-temperature transportation on the kit, is stable and efficient, and can meet various requirements of enzyme-linked immunoassay kit for chromogenic substrate solutions.

Description

technical field [0001] The invention relates to the technical field of clinical in vitro detection, in particular to a stable and efficient HRP enzymatic substrate solution and its preparation method and application. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) is a technique in which known antigens or antibodies are adsorbed on the surface of a solid phase carrier, and the enzyme-labeled antigens and antibodies are reacted on the solid phase surface. This technology can be used to detect macromolecular antigens and specific antibodies, etc., and has the advantages of rapidity, sensitivity, simplicity, and easy standardization of carriers. In 1971, Engvall and Perlmann published an article on the use of enzyme-linked immunosorbent assay (Enzyme linked immunosorbent assay, ELISA) for the quantitative determination of IgG, which made the enzyme-labeled antibody technology for antigen localization developed in 1966 into the detection of trace substanc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531
CPCG01N33/531
Inventor 王玲邵豪锋刘瑞鑫刘爽霍晓飞李金雨
Owner BEIJING BEIER BIOENG
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