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ELISA (enzyme-linked immunosorbent assay) plate coating liquid, confining liquid and ELISA plate manufacturing method

A production method and technology of microtiter plate, which can be used in measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to detect the uniformity of coating effectively, and achieve the effect of ensuring quality

Active Publication Date: 2016-05-18
馥申生命科学(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, in view of the technical problem that the existing pre-coated ELISA plate manufacturing method cannot effectively detect the coating uniformity, the purpose of the present invention is to provide an ELISA plate coating solution, an ELISA plate blocking solution and a A method for making a pre-coated microtiter plate that can detect coating uniformity during the coating process using the microplate coating solution and the microplate blocking solution

Method used

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  • ELISA (enzyme-linked immunosorbent assay) plate coating liquid, confining liquid and ELISA plate manufacturing method
  • ELISA (enzyme-linked immunosorbent assay) plate coating liquid, confining liquid and ELISA plate manufacturing method
  • ELISA (enzyme-linked immunosorbent assay) plate coating liquid, confining liquid and ELISA plate manufacturing method

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0029] Example 1 Preparation of Streptavidin pre-coated ELISA plate

[0030] 1.1 Reagent configuration

[0031] The reagents that need to be used in the production process include ELISA plate coating solution, washing solution and blocking solution, which can be configured according to the ingredients and parameters or contents in Table 1.

[0032] Table 1

[0033]

[0034] 1.2 Production steps

[0035] 1) Dissolve streptavidin with the ELISA plate coating solution, the concentration of streptavidin is 2μg / mL;

[0036] 2) Add streptavidin-dissolved ELISA plate coating solution (100 μL / well) to each microwell of the 96-well ELISA plate to be coated, and coat overnight at 4°C;

[0037] 3) Wash the plate 3 times with washing solution (350μL / well / time), and pat dry;

[0038] 4) Add blocking solution (250μL / well) and block at 37°C for 2 hours; use a microplate reader to detect the absorbance of the blocking solution in each microwell of the microtiter plate at a wavelength of 340nm, and calcula...

Embodiment 2

[0049] Example 2 Preparation of aflatoxin M1 monoclonal antibody pre-coated ELISA plate

[0050] 2.1 Reagent configuration

[0051] Configure the coating solution, washing solution and blocking solution of the microplate according to the ingredients and parameters or content in Table 2.

[0052] Table 2

[0053]

[0054] 2.2 Production steps

[0055] Referring to Example 1, the difference is that in step 1), streptavidin is replaced with AFM1 monoclonal antibody, and the concentration of AFM1 monoclonal antibody is 0.5 μg / mL.

[0056] 2.3 Effect test

[0057] Detection of unqualified products: A total of 50 ELISA plates were coated, of which the coefficient of variation in 7 of the wells exceeded 5%, and the unqualified rate was 14%.

[0058] Take a qualified and unqualified pre-coated ELISA plate, respectively, to detect milk samples with an AFM1 content of 0.45ppb.

[0059] The steps are:

[0060] A. Add AFM1 to milk without AFM1 to prepare a milk sample with an AFM1 content of 0.45ppb.

[...

example 3

[0068] Example 3 Preparation of aflatoxin B1 (AFB1) monoclonal antibody pre-coated ELISA plate

[0069] 3.1 Reagent preparation:

[0070] Configure the coating solution, washing solution and blocking solution for the ELISA plate according to the ingredients and parameters or content in Table 3.

[0071] table 3

[0072]

[0073] 3.2 Production steps

[0074] Referring to Example 1, the difference is that in step 1), streptavidin is replaced with AFB1 monoclonal antibody, and the concentration of AFB1 monoclonal antibody is 1 μg / mL.

[0075] 3.3 Effect test

[0076] Detection of unqualified products: A total of 50 ELISA plates were coated, of which the coefficient of variation in 2 of the wells exceeded 5%, and the unqualified rate was 4%.

[0077] Take a qualified and unqualified pre-coated ELISA plate, respectively, and use them to detect the corn flour samples with AFB1 content of 5ppb.

[0078] The steps are:

[0079] A. Take a sample of corn flour with an AFB1 content of 5ppb verified by...

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Abstract

The invention discloses an ELISA (enzyme-linked immunosorbent assay) plate coating liquid and an ELISA plate confining liquid. The ELISA plate coating liquid contains a carbonate buffer solution with pH of 9.6 and G-6-PD, and the ELISA plate confining liquid contains a phosphate buffer solution with pH of 8.0, NADP<+> and G-6-P. The invention further discloses a method for manufacturing a pre-coated ELISA plate through matching use of the ELISA plate coating liquid and the ELISA plate confining liquid. According to the method for manufacturing the pre-coated ELISA plate, G-6-PD and a to-be-coated antigen or antibody are coated together in the coating process, NADP<+> can be reduced into NADPH during closing, coating uniformity of G-6-PD can be indirectly detected through detection of NADPH, then the coating uniformity of the antigen or antibody is reflected, accordingly, an unqualified pre-coated ELISA plate is identified in a manufacturing process, and the quality of the pre-coated ELISA plate can be guaranteed.

Description

Technical field [0001] The invention belongs to the technical field of enzyme label plate coating, and specifically relates to a method for manufacturing an enzyme label plate coating solution, an enzyme label plate sealing solution and a pre-coated enzyme label plate. Background technique [0002] Enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA) has many advantages such as high sensitivity, high specificity, and easy standardization. It is widely used in biomedical in vitro diagnostics, life science research, food safety testing and other aspects. [0003] The ELISA method utilizes the specific reaction between antigen and antibody and the color reaction produced by enzyme and substrate. The principle is that the antigen or antibody is linked to the enzyme to form an enzyme-labeled antigen or antibody, and then reacts with the corresponding antibody or antigen, and then the enzyme reacts with the substrate. The color reaction of the substance can be qua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/31
CPCG01N21/31G01N33/543
Inventor 危林丹韩敏余春湖
Owner 馥申生命科学(上海)有限公司
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