Indirect ELISA (enzyme linked immunosorbent assay) method for detecting soybean antigenic protein serum antibody

A technology of soybean antigenic protein and serum antibody, which is applied in the direction of biological testing, material inspection products, etc., can solve the problems of delayed disease, economic loss, and inability to remove allergens in time, and achieve strong sensitivity and specificity, simple detection method, The effect of resolving soy protein-derived food allergy

Inactive Publication Date: 2013-01-16
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can only be used as a preventive method. It cannot effectively identify and diagnose allergic animals, delays the disease, and cannot remove allergens in time, leading to aggravation of the animal's condition and causing major economic losses.

Method used

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  • Indirect ELISA (enzyme linked immunosorbent assay) method for detecting soybean antigenic protein serum antibody
  • Indirect ELISA (enzyme linked immunosorbent assay) method for detecting soybean antigenic protein serum antibody
  • Indirect ELISA (enzyme linked immunosorbent assay) method for detecting soybean antigenic protein serum antibody

Examples

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Effect test

Embodiment example 1

[0020] Indirect ELISA method provided by the present invention, wherein, with glycinin (11S protein) as detection antigen:

[0021] (1) Antigen purification and purity analysis

[0022] Purification: 11S protein was dissolved in phosphate buffer with a pH of 7.4, Sepharose CL-6B gel filtration (1.6×120 cm chromatography column), and the eluate obtained by elution with phosphate buffer was dialyzed, Freeze-dried glycinin (ie, 11S protein) was stored at -20°C as an antigen for future use. in figure 1 It is an explanatory diagram of the protein elution peak data of glycinin (11S protein), and the purification data can be referred to figure 1 .

[0023] Purity analysis: fix the gel plate and install it in the electrophoresis tank, take 10 μL of standard (sample) protein solution into the EP tube, then add 10 μL of 2 times sample buffer, the sample volume is 10 μL. Before adding the sample, the sample was heated in boiling water for 3-5 minutes, cooled and set aside. According...

Embodiment example 2

[0040] The indirect ELISA method provided by the present invention, wherein, with β-conglycinin (7S protein) as detection antigen:

[0041] (1) Antigen purification and purity analysis

[0042] Purification: 7S protein was dissolved in phosphate buffer with a pH of 7.4, Sepharose CL-6B gel filtration (1.6×120 cm chromatography column), and the eluate obtained by elution with phosphate buffer was dialyzed, Freeze-dried to obtain β-conglycinin (ie, 7S protein), which was stored at -20°C as an antigen for future use. in, image 3 It is an explanatory diagram of the protein elution peak data of β-conglycinin (7S protein), and the purification data can be referred to image 3 .

[0043] Purity analysis: fix the gel plate and install it in the electrophoresis tank, take 10 μL of standard (sample) protein solution into the EP tube, then add 10 μL of 2 times sample buffer, the sample volume is 10 μL. Before adding the sample, the sample was heated in boiling water for 3-5 minutes,...

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Abstract

The invention discloses an indirect ELISA (enzyme linked immunosorbent assay) method for detecting soybean antigenic protein serum antibody. The indirect ELISA method includes using purified soybean antigenic protein as antigenic peridium to detect soybean antigenic protein serum antibody. The soybean antigenic protein comprises glycinin and beta-conglycinin. The indirect ELISA method has the advantages that the method has high sensitivity and specificity to allergic diseases, which are caused by the soybean antigenic protein, of humans and animals, a basis is provided for diagnosis of the diseases, soybean protein source food allergy of animals in our country is convenient to overcome, effective technical means is provided for disease research of the diseases, and the quick and simple detecting method is provided for mastering prevention and morbidity of the soybean antigenic protein allergic diseases in humans and animals.

Description

technical field [0001] The invention relates to an indirect ELISA method for detecting serum antibodies of soybean antigen protein, belonging to the technical field of biological detection. Background technique [0002] Soybean is the most widely used high-quality plant protein source in animal breeding. It not only has high protein content and low price, but also has high absolute content of essential amino acids and a balanced composition ratio of amino acids. However, the allergens present in soybeans can trigger allergic reactions in humans and animals, especially infants and young animals, leading to diarrhea, digestive and absorption disorders, and even death, causing heavy losses and limiting its use in production. application. Studies have shown that the main sensitizing factor that causes allergic reactions in animals is soybean antigenic protein, and glycinin (glycinin) and β-conglycinin (β-conglycinin) are the main antigenic components in soybean antigenic protei...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 吴金节王希春寇亚楠孙志阔徐述亮陈亮王勇马良友
Owner ANHUI AGRICULTURAL UNIVERSITY
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