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ELISA (enzyme-linked immunosorbent assay) chromogenic substrate and preparation method thereof

A chromogenic substrate and enzyme-linked immunosorbent technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high operational accuracy, short storage validity period, and difficult comparison and verification of test data, so as to reduce batch-to-batch difference, simplified detection operation steps, and excellent stability

Active Publication Date: 2014-07-09
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] At present, there are many ways to prepare chromogenic substrates. The commercially available chromogenic substrates are not only expensive, but also have a short shelf life, and require high operational accuracy during use; many laboratories prepare them by themselves, but there are various preparation methods There are no unified standards and specifications; the chromogenic substrates used in research papers are also varied, which makes it difficult for the comparison and verification of detection data

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Add 1.0 g of citric acid monohydrate and 26 g of disodium hydrogen phosphate dodecahydrate to 800 mL of distilled water, adjust the pH to 4.0 with 6M hydrochloric acid, add distilled water to make the volume to 1000 mL to prepare a buffer mother liquor; add 3.2 to the buffer mother liquor g tetramethyl benzidine hydrochloride and 0.2 g disodium edetate, stir to dissolve to obtain a mixed solution; add 15 mL of glycerol, 98 mg of sodium pyrophosphate and 9 mg of sodium stannate to the mixed solution, and protect from light Stir overnight to obtain the color-developing substrate.

Embodiment 2

[0027] Add 1.5 g of citric acid monohydrate and 30 g of disodium hydrogen phosphate dodecahydrate to 800 mL of distilled water, adjust the pH to 4.5 with 6M hydrochloric acid, add distilled water to make the volume to 1000 mL to prepare a buffer mother liquor; add 3.5 to the buffer mother liquor g tetramethylbenzidine hydrochloride and 0.3g disodium ethylenediaminetetraacetic acid, stir to dissolve to obtain a mixed solution; add 20mL glycerol, 100mg sodium pyrophosphate and 10mg sodium stannate to the mixed solution, and protect from light Stir overnight to obtain the color-developing substrate.

Embodiment 3

[0029] Add 1.2 g of citric acid monohydrate and 28 g of disodium hydrogen phosphate dodecahydrate to 800 mL of distilled water, adjust the pH to 4.2 with 6M hydrochloric acid, and add distilled water to make the volume to 1000 mL to prepare a buffer mother liquor; add 3.4 to the buffer mother liquor g tetramethylbenzidine hydrochloride and 0.25g disodium edetate, stir to dissolve to obtain a mixed solution; add 17mL glycerol, 99mg sodium pyrophosphate and 10mg sodium stannate to the mixed solution, protect from light Stir overnight to obtain the color-developing substrate.

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PUM

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Abstract

The invention relates to an ELISA (enzyme-linked immunosorbent assay) chromogenic substrate of which the pH value is 4.0-4.5. The ELISA chromogenic substrate is composed of 0.9-1.5 g / L citric acid monohydrate, 25-30 g / L disodium hydrogen phosphate dodecahydrate, 3-3.5 g / L tetramethyl benzidine hydrochloride, 0.15-0.3 g / L disodium edetate, glycerol (the volume ratio of the glycerol to the chromogenic substrate is 0.01-0.02), 0.095-0.1 g / L sodium pyrophosphate, 0.008-0.01 g / L sodium stannate, pH regulator and water. By using the tetramethyl benzidine hydrochloride as the main chromogenic component, the chromogenic substrat can be directly completely dissolved in a water solution, has high stability, and does not need mixing in advance, thereby simplifying the operation steps and reducing the differences among batches.

Description

Technical field [0001] The invention belongs to the technical field of biological in vitro diagnostic reagents, and specifically relates to an enzyme-linked immunochromogenic substrate and a preparation method thereof. Background technique [0002] In 1971, the Swedish scholars Engvail and Perlmann, and the Dutch scholars Van Weerman and Schuurs respectively reported the development of immunological technology into a solid-phase immunoassay method for detecting trace substances in body fluids, namely enzyme-linked immunosorbent assay (ELISA). ELISA has now become a frontier subject in the field of analytical chemistry. It is a special reagent analysis method and a new type of immunoassay technology developed on the basis of immunoenzymatic techniques. [0003] Fundamental: [0004] It uses the specific reaction of the antigen and the antibody to connect the test substance with the enzyme, and then the enzyme and the substrate produce a color reaction for quantitative determination. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N21/78
CPCG01N33/535
Inventor 李庆春秦枫王秀伟
Owner SUZHOU HAOOUBO BIOPHARML
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