A kind of enzyme-linked immunochromogenic substrate and preparation method thereof

A chromogenic substrate and enzyme-linked immunosorbent technology, which is applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of high operational accuracy, short storage period, difficulty in comparison and verification of test data, and achieve excellent stability. , reduce the difference between batches, and simplify the detection operation steps

Active Publication Date: 2016-01-06
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 hydrochloric acid with a concentration of 6 M, and add distilled water to make the volume to 1000 mL to prepare a buffer mother solution; add 3.2 g tetramethylbenzidine hydrochloride and 0.2 g disodium edetate were stirred and dissolved to obtain a mixed solution; 15 mL glycerol, 98 mg sodium pyrophosphate and 9 mg sodium stannate were added to the mixed solution to avoid light Stir overnight to obtain a chromogenic substrate.

Embodiment 2

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

Embodiment 3

[0029] Add 1.2g of citric acid monohydrate and 28g of disodium hydrogen phosphate dodecahydrate to 800mL of distilled water, adjust the pH to 4.2 with hydrochloric acid with a concentration of 6M, add distilled water to make the buffer mother solution to 1000mL; add 3.4 g of tetramethylbenzidine hydrochloride and 0.25 g of disodium edetate were stirred and dissolved to obtain a mixed solution; 17 mL of glycerol, 99 mg of sodium pyrophosphate and 10 mg of sodium stannate were added to the mixed solution to avoid light Stir overnight to obtain a chromogenic 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, in particular to an enzyme-linked immunochromogenic substrate and a preparation method thereof. Background technique [0002] In 1971, Swedish scholars Engvail and Perlmann, Dutch scholars VanWeerman and Schuurs respectively reported the development of immune technology into a solid-phase immunoassay method for detecting trace substances in body fluids, namely enzyme-linked immunosorbent assay (enzyme-linkedimmunosorbentassay, ELISA). ELISA has now become a frontier topic 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 antigen and antibody to link the analyte to the enzyme, and then produces a color reaction through the enzyme and the substrate for quantitative determination. T...

Claims

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Application Information

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Patent Type & Authority Patents(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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