IgE binding epitope of main allergen alpha-lactalbumin of cow milk whey

A technology for major allergens and lactalbumin, which is applied in the field of IgE binding epitopes of the major allergen α-lactalbumin in bovine milk whey, can solve the problem of less reporting of bovine milk whey allergen epitope localization

Inactive Publication Date: 2022-07-22
谭宏凯
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many epitope mapping studies have been carried out on milk allergens, including casein, β-lactoglobulin, and α-lactalbumin, there are few reports on the epitope mapping of bovine milk whey allergens specific to the Chinese population

Method used

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  • IgE binding epitope of main allergen alpha-lactalbumin of cow milk whey
  • IgE binding epitope of main allergen alpha-lactalbumin of cow milk whey
  • IgE binding epitope of main allergen alpha-lactalbumin of cow milk whey

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0019] 1. Construction of serum pools for patients with milk allergy

[0020] 1. Indirect ELISA to screen serum of patients with milk allergy

[0021] The level of sIgE against bovine milk protein in the serum of allergic patients was detected by ImmunoCAP, and sera with bovine milk protein sIgE level greater than or equal to 0.35kUA / L and aged 3–12 years were selected for ELISA. Determination of anti-α-lactalbumin specific immunoglobulin E (ALA-sIgE) in serum of patients with cow's milk allergy by indirect ELISA method (for the indirect ELISA method, please refer to Li Liuxu's paper, the source of the paper: Li Liuxu. Based on ELISA method Study on the linear epitope of ovomucoid IgE [D]. Tianjin Medical University, 2020: 11-16.).

[0022] The coating solution is 0.05mol / L carbonate buffer with pH 9.6. The washing solution, blocking solution, serum, secondary antibody, etc. are all diluted with 0.01M PBS buffer. The washing solution contains 0.05% (v / v) Tween-20, blocking s...

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Abstract

The invention relates to an IgE (Immunoglobulin E) binding epitope of a whey allergen alpha-lactalbumin. The amino acid sequence of the IgE binding epitope is 11Glu-Leu-Lys-Asp-Leu-Lys-Gly-Tyr-Gly-Gly-Val-Ser-Leu-Pro-Glu-Trp-Val 27. The invention further relates to a preparation method of the IgE binding epitope. According to the preparation method disclosed by the invention, 16 parts of serum are screened by adopting an enzyme-linked immunosorbent assay (ELISA), and finally 12 parts of serum of a child patient with cow milk allergy are obtained. High-purity cow milk specific IgE purified by a Hitrap Protein G HP affinity chromatography column is used as a target molecule, and affinity panning is carried out on a phage dodecapeptide library. And predicting the linear epitope by using bioinformatics software, and finally obtaining the IgE epitope information of the whey allergen alpha-lactalbumin. The invention has great significance for ascertaining the immunological mechanism of cow milk allergy, researching the effect of the epitope in food anaphylaxis and guiding the development of hypoallergenic dairy products for directionally breaking the epitope.

Description

technical field [0001] The invention relates to the fields of molecular biology, immunology and bioinformatics, in particular to the IgE binding epitope of alpha-lactalbumin, the main allergen of bovine whey. Background technique [0002] Milk and dairy products are favored by consumers because of their high nutritional value, but food safety issues caused by milk allergy have also attracted widespread attention. Milk and milk products are one of the eight major allergic foods identified by WHO / FAO. Cow's milk allergy (CMA) is a common disease early in life with a prevalence of 2–7% and an increasing trend. It is estimated that 2–3% of infants in European countries develop symptoms of cow's milk allergy. In a recent survey of 6768 Chinese infants, the prevalence of CMA was found to be 2.69%. CMA is divided into immunoglobulin E (immunoglobulin E, IgE) mediated, non-IgE mediated and mixed (IgE and non-IgE) mediated. About 60% of CMAs are IgE-mediated, i.e., associated wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/76
CPCC07K14/76
Inventor 谭宏凯胡巍熊子奕胡永芯邱毓黎晶晶李欣陈红兵武涌孟轩夷
Owner 谭宏凯
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