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Detection of animal-derived allergen bovine serum albumin by using indirect competitive ELISA method

A technology for serum albumin and animal detection, which is applied in the field of analysis and detection, and can solve the problems of less reports of allergen BSA detection methods and the like

Inactive Publication Date: 2017-02-15
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports on the detection methods for the allergen BSA at present.

Method used

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  • Detection of animal-derived allergen bovine serum albumin by using indirect competitive ELISA method
  • Detection of animal-derived allergen bovine serum albumin by using indirect competitive ELISA method
  • Detection of animal-derived allergen bovine serum albumin by using indirect competitive ELISA method

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Experimental program
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Embodiment Construction

[0037] The present invention will be further described below in conjunction with embodiment, but protection scope of the present invention is not limited to this:

[0038] cells and experimental animals

[0039] Mouse myeloma cells SP2 / 0 were purchased from Shanghai Cell Institute, Chinese Academy of Sciences. BALB / c female mice aged 6-8 weeks were purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences.

[0040] main reagent

[0041] Bovine serum albumin (BSA) and polyethylene glycol (PEG) were purchased from Sigma, USA; protein quantification kit (BCAProtein Assay kit), Novagen, USA; Freund's complete adjuvant (FCA), Freund's incomplete adjuvant Chromogenic solution (FIA) and tetramethylbenzidine (TMB) were purchased from Promega, USA; goat anti-mouse IgG horseradish peroxide (HRP) labeled antibody was purchased from Proteintech, USA; polyvinylidene fluoride membrane ( PVDF) was purchased from Miliipore, USA. Major equipment

[0042] Micropipette...

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Abstract

The invention discloses detection of animal-derived allergen bovine serum albumin by using an indirect competitive ELISA method, and belongs to the field of analysis detection. According to the method, BSA is used as a coating antigen and a standard substance, high-specificity and high-purity anti-BSA monoclonal antibody is screened as primary antibody, goat anti-mouse IgG horseradish peroxidase (HRP) labeled antibody is adopted as secondary antibody, tetramethyl benzidine (TMB) is adopted as a substrate, sensitive and specific indirect competitive ELISA is established, and the results are analyzed by using a mathematic method. The method of the present invention has advantages of strong specificity, high sensitivity and good repeatability, and can be used for rapid detection of allergen BSA.

Description

technical field [0001] The invention belongs to the field of analysis and detection, and in particular relates to an indirect competitive ELISA method for detecting animal-derived allergen bovine serum albumin. Background technique [0002] Food allergy is an immune-mediated adverse reaction of the human body to antigenic substances in food, which triggers a series of clinical pathophysiological changes in the body. In severe cases, anaphylactic shock may occur, and even life-threatening. According to the statistics of the World Health Organization (WHO), about 25% of people in the world suffer from allergic diseases, and the rate of increase is 23 times every 10 years. There are more than 200 million patients with food allergens in my country. In recent years, with the acceleration of industrialization, urbanization, and globalization, people's life rhythms and styles are constantly changing, life pressure is increasing, and there are more and more food types and processing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6803
Inventor 王玮朱业培吕青骎徐幸莲周光宏
Owner NANJING AGRICULTURAL UNIVERSITY