Method for assisting in identifying linear epitope of plant food allergen

A food allergy, plant-based technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of high blindness, heavy workload, long time consumption, etc., and achieve the effect of accurate identification results and reduced workload.

Active Publication Date: 2020-05-01
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The peptide synthesis technology is to incubate the synthesized short overlapping peptides with the serum of allergic patients through dot blotting or immunoblotting to detect their binding to specific IgE antibodies, thereby judging and screening linear epitopes, but it is necessary to collect serum from allergic patients , time-consuming, heavy workload, high blindness, it is possible to ignore epitopes in overlapping intervals, and it is difficult to guarantee accuracy

Method used

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  • Method for assisting in identifying linear epitope of plant food allergen
  • Method for assisting in identifying linear epitope of plant food allergen
  • Method for assisting in identifying linear epitope of plant food allergen

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Peanut allergen Ara h1 is a 7S globulin, which belongs to the Cupin superfamily, and can be recognized by more than 90% of peanut-allergic patients' sera through serological analysis. Because the structural sequence of the Cupin superfamily has the characteristics of high conservation, strong thermal stability, resistance to enzymolysis and indigestibility, Ara h1 is selected as the model allergen in the present invention.

[0035] This embodiment provides a method for assisting in identifying the linear epitope of the peanut allergen Ara h1, comprising the following steps:

[0036] S1. The product of peanut protein digested by pepsin for 60 minutes was used as the sample, and the nano-level HPLC liquid phase system Easy-nLC1000 was used for separation, wherein the buffer solution: solution A was 0.1% formic acid aqueous solution, and solution B was 0.1% formic acid acetonitrile solution; The chromatographic column is balanced with 95% liquid A; the sample is loaded fro...

Embodiment 2

[0050] The walnut allergen Jug r 2 belongs to the 7S vicilin family, which is the same family as the peanut allergen Ara h1. Its primary sequence consists of 593 amino acids and its molecular weight is about 44kDa. It exists in the form of trimeric oligomers under natural conditions.

[0051] This embodiment provides a method for assisting in identifying the linear epitope of the walnut allergen Jug r 2, comprising the following steps:

[0052] S1. The product of walnut protein digested by pepsin for 60 minutes was used as a sample, and the nano-level HPLC liquid phase system Easy-nLC1000 was used for separation, wherein the buffer solution: solution A was 0.1% formic acid aqueous solution, and solution B was 0.1% formic acid acetonitrile solution; The chromatographic column is balanced with 95% liquid A; the sample is loaded from the autosampler to the pre-column C18trap column (3mm, 0.10×20mm), and then separated by the analytical column C18column (1.9mm, 0.15×120mm), with a ...

Embodiment 3

[0067] Wheat allergen CM16 is a protein with a molecular weight of 17kDa and composed of 143 amino acids. It belongs to the gliadin superfamily and contains multiple cysteine ​​residues, which can form intramolecular disulfide bonds to ensure the structural stability of the protein and It is heat resistant.

[0068] This embodiment provides a method for assisting in identifying the linear epitope of wheat allergen CM16, comprising the following steps:

[0069] Using the product of wheat protein digested by trypsin for 60 minutes as a sample, a total of 6753 peptides were detected by using the steps S1-S3 in Example 1, and a search database was constructed according to the amino acid sequence of wheat allergens provided by the Uniprot database, and searched with pFind The engine detected a total of 23 peptides belonging to wheat allergen proteins, most of which were derived from α-amylase / trypsin inhibitor subtypes and high molecular weight glutenin subunits, and 3 of them matc...

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Abstract

The invention provides a method for assisting in identifying linear epitope of a plant food allergen. The method comprises the following steps of taking a product of a plant food protein which is subjected to simulating gastrointestinal digestion as a sample, and carrying out high-performance liquid chromatography; carrying out mass spectrum detection on the separated sample to obtain mass spectrum information of an anti-digestion peptide segment; and next, performing comparison and analysis with an amino acid full sequence of the plant food allergen to obtain the peptide segment matched withthe plant food allergen. The peptide segment obtained by the method of the invention has correlation, or coverage or cross with the linear epitope of the allergen. Therefore, the range for identifyingthe linear epitope can be reduced and the identifying can be more targeted, and compared with the existing method for identifying the linear epitope, the assisting method reduces the workload of thewhole identification process, and the identification result is more accurate.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to a method for assisting in identifying linear epitopes of plant food allergens. Background technique [0002] Food allergy is an adverse reaction that occurs when the body is exposed to a specific food, that is, an allergic reaction that may involve different organs of the body caused by the immune system's specific immune response to the allergen. According to epidemiological surveys, the incidence rate of infants and young children worldwide has reached 8%, and that of adults has reached 5%. Most food allergies are type I hypersensitivity reactions mediated by IgE, which can cause systemic multi-system reactions such as asthma, diarrhea, urticaria, and allergic dermatitis, and even cause life-threatening shock and seriously affect the quality of life of patients. [0003] There are many kinds of food allergens in nature, mainly derived from plants, animals and mic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 车会莲杨帅李欣芮陈成范卓妍张亚妮
Owner CHINA AGRI UNIV
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