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Antigen region for processing destroyed beta-conglycinin alpha'subunit based on phage showing technology positioning and screening method thereof

A conglycinin and phage display technology, which is applied in the fields of immunology, bioinformatics, and molecular biology, can solve the problems of no relevant literature on allergen epitopes

Inactive Publication Date: 2018-11-02
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the β-conglycinin α' subunit, and the application of phage display technology to the epitope mapping of the soybean allergen β-conglycinin α' subunit has not been reported, especially the localization by The allergen epitopes destroyed after processing have not been found in the relevant literature

Method used

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  • Antigen region for processing destroyed beta-conglycinin alpha'subunit based on phage showing technology positioning and screening method thereof
  • Antigen region for processing destroyed beta-conglycinin alpha'subunit based on phage showing technology positioning and screening method thereof
  • Antigen region for processing destroyed beta-conglycinin alpha'subunit based on phage showing technology positioning and screening method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Preparation of allergen epitope specific antibody destroyed by ultra-high pressure treatment

[0029] 1. Ultra-high pressure treatment of β-conglycinin:

[0030] Put β-conglycinin with a concentration of 15 mg / mL in a sterile homogenization bag, seal and vacuum, place the sealed homogenization bag in the processing chamber (23°C) of the ultra-high pressure processing device, and start When the pressure is increased to 455MPa, the pressure is increased to 455MPa, and the pressure is maintained for 18 minutes, and then the pressure is relieved. The pressure relief rate is 300 MPa / min. The inhibitory rate of β-conglycinin antigen after treatment was 49.59%.

[0031] 2. Preparation of allergen epitope-specific antibodies destroyed by ultra-high pressure treatment (rabbit antiserum absorbed by the ultra-high pressure treatment antigen):

[0032] New Zealand white rabbits were fed with natural β-conglycinin to prepare β-conglycinin polyclonal antibody (rabbit antiserum). ...

Embodiment 2

[0033] Example 2. Preparation of specific antibody against allergen epitope destroyed by heat treatment

[0034] 1. Heat treatment of β-conglycinin:

[0035] Place β-conglycinin at a concentration of 10 mg / mL in a beaker and heat in a water bath at 90°C for 60 minutes. The inhibitory rate of β-conglycinin antigen after treatment was 38.4%.

[0036] 2. Preparation of heat-treated allergen epitope specific antibody (rabbit antiserum absorbed by heat-processed antigen):

[0037] The method is the same as in Example 1.

Embodiment 3

[0038] Example 3. Sectional cloning of β-conglycinin α'subunit gene overlapping (overlapping)

[0039] 1. Cloning of β-conglycinin α'subunit gene

[0040] The amino acid sequence of the α'subunit of β-conglycinin was retrieved through NCBI GenBank database. Log in to the PDB database to search for the homologous protein of the α'subunit of β-conglycinin as a prediction template for the tertiary structure. Log in to the SWISS-MODEL homology modeling server to predict the tertiary structure of β-conglycinin α'subunit. According to the tertiary structure model of β-conglycinin α'subunit, its conformational epitope is predicted in the DiscoTope 2.0 web server. According to the predicted position of the tertiary structure of the protein and the B cell conformational epitope, the β-conglycinin α'subunit amino acid sequence is divided into 3 segments, and each sequence overlaps the previous sequence by about 10-50 amino acid residues (underlined Marked). The design of overlapping frag...

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Abstract

The invention discloses an antigen region for processing destroyed beta-conglycinin alpha'subunit based on phage showing technology positioning and a screening method thereof. The amino acid sequenceof the antigen region is shown as SEQ ID NO.11. The antigen region for processing destroyed beta-conglycinin is researched by using a series of bioinformatics software, referring the beta-conglycininthree-dimensional crystalline structure analyzed by a PDB database and using a phage showing technology; the total length of the beta-conglycinin alpha'subunit and overlapping segmented protein are shown on the phage surface, so that regions with beta-conglycinin alpha'subunit antigenicity reduction caused by different processing methods are precisely positioned. Theoretical basis of processing method screening is provided for food industry; an application product for fast detecting the processed food desensitization effect can be further developed.

Description

Technical field [0001] The present invention relates to the fields of molecular biology, immunology, and bioinformatics, in particular to a processing and destruction of β-conglycinin α'subunit antigen region based on phage display technology positioning. Background technique [0002] Allergic reaction is also called allergy or hypersensitivity reaction, which is a common reaction in human autoimmune diseases. Food allergies account for 50% of all allergic diseases. Soy is one of the eight major food allergens. Soybeans and their soybean products, which existed as early as 2000 BC, have long appeared in Asian diets, and the soybeans, which are rich in protein, oil and other functional substances, are used as a good source of nutrients. And Latin America, Africa and India also use the more common soybeans as one of the basic foods. According to investigations and studies, the sensitization rate of soybeans in the general population is about 0.5%, while in children it is as high...

Claims

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Application Information

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IPC IPC(8): C07K14/415
CPCC07K14/415
Inventor 席俊李爽皮江一陈珍妮刘德果
Owner HENAN UNIVERSITY OF TECHNOLOGY
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