Positioned processed and damaged Gly m Bd 60K protein antigen region based on phage display technology and screening method

A phage display and protein technology, applied in the fields of molecular biology, bioinformatics, and immunology, which can solve problems such as no related literature.

Inactive Publication Date: 2018-07-27
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on soybean allergen Gly m Bd 60K protein mainly focuses on the allergenicity identification of Gly m Bd 60K protein, linear epitope prediction and positioning, detection analysis, desensitization technology research, etc., and the application of phage display technology to soybean allergen The research on the epitope localization of Gly m Bd 60K protein has not been reported yet, especially the localization of allergen epitopes destroyed after processing has not been found in the relevant literature

Method used

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  • Positioned processed and damaged Gly m Bd 60K protein antigen region based on phage display technology and screening method
  • Positioned processed and damaged Gly m Bd 60K protein antigen region based on phage display technology and screening method
  • Positioned processed and damaged Gly m Bd 60K protein antigen region based on phage display technology and screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

[0030] Put β-conglycinin with a concentration of 15mg / mL in a sterile homogeneous bag, seal the bag and vacuumize it, and place the sealed homogeneous bag in the processing chamber (23°C) of the ultra-high static pressure processing device , start boosting, the boosting rate is 250MPa / min, when the pressure rises to 455MPa, keep the pressure for 18min, and then release the pressure, the pressure releasing rate is 300MPa / min. The inhibition rate of β-conglycinin antigen after treatment was 49.59%.

[0031] 2. Preparation of allergen epitope-specific antibodies (antigen-absorbing polyclonal antibodies) destroyed by ultrastatic high-pressure treatment:

[0032] New Zealand white rabbits were fed with natural β-conglycinin to prepare polyclonal antibody to β-conglycinin. Add excess β-conglyci...

Embodiment 2

[0033] Example 2, Preparation of Allergen Epitope-Specific Antibodies Destroyed by Heat Treatment

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

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

[0036] 2. Preparation of allergen epitope-specific antibodies destroyed by heat treatment:

[0037] Method is with embodiment 1.

Embodiment 3

[0038] Example 3. Preparation of Allergen Epitope-Specific Antibodies Destroyed by Glycosylation Treatment

[0039] 1. Glycosylation treatment of β-conglycinin:

[0040] Dissolve β-conglycinin and glucose (mass ratio: 4:1) in distilled water to make the final concentration of the mixed solution 6%, mix well and vacuum freeze-dry, and place the freeze-dried sample at constant temperature and humidity In the incubator, adjust the relative humidity to 79%, and the temperature to 60° C., and react for 2.5 days. The inhibition rate of β-conglycinin antigen after treatment was 42.8%.

[0041] 2. Preparation of allergen epitope-specific antibodies destroyed by glycosylation:

[0042] Method is with embodiment 1.

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Abstract

The invention discloses a positioned processed and damaged Gly m Bd 60K protein antigen region based on a phage display technology and a screening method. An amino acid sequence of the antigen regionis shown in SEQ ID NO.16. The method utilizes a series of bioinformatics software and refers to a three-dimensional crystal structure of analyzed beta-conglycinin in a PDB database, studies the processed and damaged Gly m Bd 60K protein antigen region by the phage display technology, and performs accurate positioning on the region where antigenicity of a Gly m Bd 60K protein is reduced due to different processing methods through presenting the Gly m Bd 60K protein and an overlapping protein of the Gly m Bd 60K protein on the surface of a phage. The method provided by the invention provides a theoretical basis for processing method screening in the food industry, and an application product for rapidly detecting a desensitization effect of processed food can be further developed.

Description

technical field [0001] The present invention relates to the fields of molecular biology, immunology and bioinformatics, in particular to an antigenic region of processing and destroying Gly m Bd 60K protein positioned based on phage display technology. Background technique [0002] Soybean is native to China and has been planted for nearly 7,000 years. The protein content in soybean is 35%-38%, and it is rich in unsaturated fatty acids, vitamins, minerals and other nutrients. It has extremely high nutritional value and is an important source of high-quality protein. It has been widely used in food processing. However, a small number of people cannot benefit from it because they are allergic to soybeans and its products. The usual symptoms are rash, itchy skin, and diarrhea. At the same time, surveys have shown that with the increase in the consumption of soybean products, the incidence of soybean allergy continues to rise. In addition, soybean allergen protein can also cau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/70G01N33/68
CPCC07K14/415C12N15/70G01N33/68G01N2333/415
Inventor 席俊贺梦雪谢岩黎于秋荣陈阳
Owner HENAN UNIVERSITY OF TECHNOLOGY
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