Method for fixing antibody in solid phase carrier in immune adsorption reaction process

A technology of solid-phase carrier and reaction process, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of reduced sensitivity, space occupation, and waste of antibodies, so as to reduce the steric hindrance of the reaction space and reduce the solid-phase concentration , the effect of reducing production costs

Inactive Publication Date: 2013-02-06
伊艾博(武汉)科技股份有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Because the physical adsorption process is relatively simple and random, and the binding sites with the carrier are diversified; and the antibody is blocked under the inert protein, there is a certain steric hindrance in the antigen-antibody binding reaction during the detection process, which affects the immunodetection

Method used

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  • Method for fixing antibody in solid phase carrier in immune adsorption reaction process
  • Method for fixing antibody in solid phase carrier in immune adsorption reaction process
  • Method for fixing antibody in solid phase carrier in immune adsorption reaction process

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Embodiment Construction

[0024] Due to the different selection of solid-phase carriers in this method, the final selected experimental steps are different. The method of the present invention will be described in detail below by taking the solid-phase carrier as a polystyrene microplate plate as an example.

[0025] 1. Materials and reagents:

[0026] 1) Antigen and antibody

[0027] Antibody: rabbit anti-FITC antibody, FITC-goat anti-human REG-4, HRP-goat anti-human REG-4

[0028] Antigen: human REG-4 standard

[0029] 2) Solid phase carrier: 96-well polystyrene microtiter plate

[0030] 3) Reagents

[0031] Coating liquid: molar concentration is 0.05mol / L, pH=9.5 sodium carbonate-sodium bicarbonate buffer solution (CBS), molar concentration is 0.05mol / L, pH=9.5 sodium carbonate-sodium bicarbonate buffer solution prepares 1.59g Sodium carbonate and 2.93g of sodium bicarbonate were dissolved in double distilled water to 1000mL, and the pH was adjusted to 9.5 with hydrochloric acid or sodium hydrox...

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Abstract

The invention relates to a method for fixing an antibody in a solid phase carrier in the immune adsorption reaction process. The method is characterized by comprising the following steps: enveloping FITC (fluorescein isothiocyanate) antibody on the solid phase carrier in advance, and then reacting 8-12 hours at 4-6 DEG C; sealing unreacted sites of the solid phase carrier, and then reacting 1-2 hours at 34-37 DEG C; adding an FITC mark antibody, then reacting 1-2 hours at 34-37 DEG C or 8-12 hours at 4-6 DEG C; and finally completing the enveloping of the solid phase carrier. According to the method provided by the invention, the problems that a current enveloped antibody is high in concentration, low in sensitivity, and poor in specificity are solved. The method that an enveloping layer is formed on the solid phase carrier in advance, and then the antibody is fixed has the advantages of simplicity, low cost, strong applicability, wide range and the like.

Description

technical field [0001] The invention relates to a method for immobilizing antibodies on a solid-phase carrier during an immunoadsorption reaction. Background technique [0002] The first condition in the process of immunoadsorption reaction is to immobilize the target antibody on the reaction carrier. The traditional method is that the antibody is immobilized on the carrier by physical adsorption, that is, the antibody is passively adsorbed on the surface of the hydrophobic solid phase carrier by physical force, and then other adsorption sites of the ELISA carrier are blocked by inert proteins. Because the physical adsorption process is relatively simple and random, and the binding sites with the carrier are diversified; and the antibody is blocked under the inert protein, there is a certain steric hindrance in the antigen-antibody binding reaction during the detection process, which affects the immunodetection Reagent sensitivity. And the antibody is distributed unevenly,...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 齐兵玉李学斌朱峰胜
Owner 伊艾博(武汉)科技股份有限公司
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