Double antibody sandwich enzyme-linked immuno sorbent assay (ELISA) method for detecting peanut allergic component Arah1

A double-antibody sandwich and allergy technology, applied in the field of immune analysis, can solve the problems of high cost, slow detection speed, and restrictions on the development of allergen detection methods, and achieve high accuracy, high precision, and high specificity

Inactive Publication Date: 2012-09-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many methods for allergen detection at present, they all have different problems, such as slow detection speed, hi

Method used

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  • Double antibody sandwich enzyme-linked immuno sorbent assay (ELISA) method for detecting peanut allergic component Arah1
  • Double antibody sandwich enzyme-linked immuno sorbent assay (ELISA) method for detecting peanut allergic component Arah1

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Embodiment Construction

[0029] specific implementation plan

[0030] The present invention is further illustrated by the following examples.

[0031] one, instrument:

[0032] TGL-40B desktop low-speed centrifuge, Shanghai Anting Scientific Instrument Factory

[0033] KFLOW pure water machine, KFLOW company

[0034] ZD-9556 horizontal shaker, Taicang Science and Education Equipment Factory

[0035] Costar 96-well 8×12 detachable microtiter plate, Shanghai Jitai Biotechnology Co., Ltd.

[0036] MuLtiska Mks Microplate Reader, Thermo Labsystems

[0037] Adjustable pipette, Thermo Labsystems

[0038] Vortex mixer, Shanghai Huxi Instrument Analysis Factory

[0039] AKTA purifier10 automatic rapid protein purification system, superdex 7510 / 300 GL gel filtration column GE health life sciences company

[0040] Vertical Electrophoresis Instrument Bio-rad Corporation

[0041] Inverted Biological Microscope Leica AG

[0042] 2. Reagents:

[0043] Horseradish peroxidase-labeled goat anti-rabbit IgG ...

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Abstract

The invention relates to a double antibody sandwich enzyme-linked immuno sorbent assay (ELISA) method for detecting a peanut allergic component Arah1, which belongs to the technical field of immunoassay. The method comprises the steps of firstly acquiring peanut protein with higher purity containing Arah1 from fresh peanut kernels, utilizing the peanut protein to immunize a mouse to acquire 14 strains of monoclonal antibodies, and selecting a combination of the monoclonal antibodies with maximal P/N value as an optimum combination. The antibody and an enzyme-labeled antibody, which are connected onto a solid-phase carrier, are respectively combined with two antigenic determinants on an antigen molecule to be detected in a sample to form a solid antibody-antigen-enzyme-labeled antibody immune compound. Since the quantity of the solid-phase antibody and the enzyme-labeled antibody is excessive relative to the antigen to be detected in the reaction system, the formation quantity of the compound is in proportion to the content of the antigen to be detected. The content of the antigen to be detected can be determined by determining the color matter amount (OD value) generated when the enzyme in the compound acts on the added substrate. The double antibody sandwich ELISA method is direct and quick, has high sensitivity, high specificity, high accuracy and high precision, is simple to operate and is used for detecting the peanut allergic component Arah1.

Description

technical field [0001] The invention relates to the preparation of a monoclonal antibody and the establishment of a double-antibody sandwich ELISA method for detecting Arah1, a peanut allergy component, and belongs to the technical field of immune analysis. Background technique [0002] In recent years, food allergy has become a public food safety problem, and peanut is one of the eight most common food allergens. Allergic diseases seriously endanger people's health. Although the minimum amount of allergens that trigger allergic reactions varies with different groups of people, a small amount of allergens can cause allergic symptoms in most patients. In order to avoid exposure to trace amounts of allergens, the detection of allergens has become a top priority. Now, although there are many allergen detection methods for reference, there are different problems in specific applications. The in vivo experimental method can provide the most direct evidence, but due to safety fa...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 匡华彭娟胥传来王利兵尹玉云宋珊珊朱建平王文彬
Owner JIANGNAN UNIV
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