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Enzyme-linked immuno sorbent assay (ELISA) enzyme immunoassay color developing agent for increasing developing duration and preparation method and application thereof

A technology of duration and color developing agent, which is applied in the direction of material inspection products, measuring devices, instruments, etc., and can solve the problems of unstable color development, color fading in the color development stage, and unstable color development process, etc.

Active Publication Date: 2014-06-25
JIANGSU BIOPERFECTUS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The addition of the reducing agent can effectively solve the problem of the precipitation of the chromogenic agent, but at the same time, another problem arises: unstable color development
When the concentration of the added reducing agent is high, the color of the product will fade rapidly after termination. Even if the concentration of the added reducing agent is very low, the color will fade slowly during the color development stage, making the entire color development process unstable, and the OD value decreases slowly The dynamic change process, which will have a negative impact on the accuracy of ELISA results

Method used

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  • Enzyme-linked immuno sorbent assay (ELISA) enzyme immunoassay color developing agent for increasing developing duration and preparation method and application thereof
  • Enzyme-linked immuno sorbent assay (ELISA) enzyme immunoassay color developing agent for increasing developing duration and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Preparation of ELISA 3,3',5,5'-tetramethylbenzidine (TMB) chromogen (1L) for increasing color duration

[0032] (1) Prepare 0.05M citric acid-sodium acetate buffer solution: Weigh 4.1g of sodium acetate and dissolve it in 1L of pure water. Then adjust the pH value of the solution with citric acid to be 5.4;

[0033] (2) Take 900mL of citric acid-sodium acetate buffer solution, put it in an ice bath, and let it stand until the system temperature reaches 4°C;

[0034] (3) Move the reaction system to a space completely protected from light, take 0.5g of TMB, dissolve it in 50mL DMSO first, and after the dissolution is complete, add it to the pre-cooled citric acid-sodium acetate buffer solution and place it in an ice bath;

[0035] (4) Add 33mL of 30% hydrogen peroxide solution, stir and mix well, add 0.05M citric acid-sodium acetate buffer solution to make the volume to 1L, and the final concentration of the substrate TMB is 0.5g / L;

[0036] (5) Take out the p...

Embodiment 2

[0039] Example 2 Preparation of ELISA enzyme immunoassay 4-chloro-1-naphthol (4C1N) chromogen (1L) for increasing color duration

[0040] (1) Prepare 0.05M citric acid-sodium acetate buffer solution: Weigh 4.1g of sodium acetate and dissolve it in 1L of pure water. Adjust the pH of the solution to 5.4 with citric acid

[0041] (2) Take 900mL of citric acid-sodium acetate buffer solution, put it in an ice bath, and let it stand until the system temperature reaches 4°C;

[0042](3) Move the reaction system to a space completely protected from light, take 1.3g of 4-chloro-1 naphthol, dissolve it in 50mL DMF first, and add it to the pre-cooled citric acid-sodium acetate buffer after the dissolution is complete. Ice bath;

[0043] (4) Add 50mL of 30% hydrogen peroxide solution, stir well and add 0.05M citric acid-sodium acetate buffer solution to make the volume to 1L. The final concentration of the substrate 4-chloro-1-naphthol is 1.3g / L ;

[0044] (5) Take out the prepared su...

Embodiment 3

[0047] Example 3 Preparation of ELISA enzyme-free nitro blue tetrazolium chloride (NBT) chromogen (1L) for increasing color duration

[0048] (1) Prepare 0.1M imidazole-HCl buffer solution: take 6.8g of imidazole hydrochloride, dissolve it in 1L of pure water, and adjust the pH to 5.4 with concentrated hydrochloric acid;

[0049] (2) Take 900mL of imidazole-HCl buffer solution, put it in an ice bath, and let it stand until the system temperature reaches 4°C;

[0050] (3) Move the reaction system to a space completely protected from light, take 5.6g of nitro blue tetrazolium chloride, dissolve it in 50mL of DMF first, and after the dissolution is complete, add it to the pre-cooled imidazole-HCl buffer solution, Ice bath;

[0051] (4) Add 50mL of 30% hydrogen peroxide solution, stir well and add 0.1M imidazole-HCl buffer solution to make the volume to 1L. The final concentration of the substrate nitro blue tetrazolium chloride (NBT) is 5.6g / L;

[0052] (5) Take out the prepa...

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Abstract

The invention discloses an enzyme-linked immuno sorbent assay (ELISA) enzyme immunoassay color developing agent for increasing developing duration and a preparation method and application thereof. Compared with the prior art, the color developing agent prepared by the method can be maintained to be close to a constant color developing state after termination within 120 minutes, so that the color developing agent maintains a quite steady state after developing is terminated. Therefore, the stability, accuracy and precision of OD value measurement are increased, and an adverse effect of the instable color developing process on the accuracy of the ELISA testing result is effectively reduced.

Description

technical field [0001] The present invention relates to an ELISA enzyme-immune chromogenic reagent for increasing the duration of color development and its preparation method and application. Using the ELISA enzyme-free chromogenic reagent of the present invention can make the color development degree of the ELISA color reaction terminated in a longer period of time. remain stable over time. Background technique [0002] The Chinese name of ELISA is "enzyme-linked immunosorbent assay", which is an immunological analysis and determination technology based on antigen-antibody reaction. It can qualitatively and quantitatively measure the antibodies of various pathogens, including IgG, IgM, IgA, etc., can be detected by this method, so as to assist clinical diagnosis of whether the subject is infected with a certain disease. This technology is widely used in medical testing, scientific research, environmental monitoring, Food safety has a wide range of applications. [0003] T...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N33/531
CPCG01N33/54393
Inventor 李荣宇段江波王国强刘中华施伟张旭
Owner JIANGSU BIOPERFECTUS TECH CO LTD
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