Method for identifying allergens

A technology of allergens and dendritic cells, applied in the field of identification of allergens, to achieve simple and effective results

Pending Publication Date: 2022-04-12
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, commonly used allergen identification methods are still mainly aimed at the effector cells of allergic reactions (mast cells and basophils), completely ignoring the influence of DC cells and T cell differentiation on food allergic reactions. blank

Method used

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  • Method for identifying allergens
  • Method for identifying allergens
  • Method for identifying allergens

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0034] (1) Isolation, purification and identification of turbot parvalbumin:

[0035] Use extraction buffer (Tris 0.1mol L -1 , Glycine 0.5mmol·L -1 , DTT 0.1mmol·L -1 ) of turbot loin was crudely extracted, and the crude extract was collected and dialyzed overnight. The dialysate was boiled in water for 5 minutes, and the supernatant was collected by centrifugation. Ammonium sulfate is added to the supernatant, so that the final concentration of ammonium sulfate in the system is 60% and 100%, and a purified turbot parvalbumin extract is obtained. Protein concentration was determined by BCA method. The protein components extracted from different extracts were separated and identified by SDS-PAGE. The electrophoretic separation gel concentration was 12% (w / v), and the loading gel concentration was 5% (w / v).

[0036] The IgG and IgE binding abilities of the purified PV were detected by Western blot, respectively. Protein bands were transferred to PVDF membranes (room temp...

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Abstract

The invention belongs to the technical field of allergen identification, and provides a method for identifying allergens. A sample to be identified is used for stimulating immature dendritic cells, flow cytometry is adopted for detecting expression of cell surface markers (MHC II and CD86), and an ELISA method is adopted for detecting the generation condition of cell factors (IL-6, IFN-gamma and TNF-alpha). Meanwhile, whether the sample to be identified is the allergen or not is judged according to the effect of the allergen to be identified on T cell differentiation after the dendritic cells are stimulated in vitro. The method disclosed by the invention is simple and effective, and provides a reference basis for rapidly identifying allergens.

Description

technical field [0001] The invention relates to the technical field of allergen identification, in particular to a method for identifying allergens. Background technique [0002] Food allergy is an adverse reaction of the human immune system to some food ingredients, and has become a food safety issue of global concern. More than 40% of the world's population suffers from food allergies. Food allergy symptoms include skin itching, vomiting, diarrhea and other gastrointestinal symptoms, which can lead to shock or even death in severe cases. With the improvement of people's living standards and the popularization of international food trade, there is an urgent need for risk assessment and mechanism exploration of food allergens. [0003] In the food-induced sensitization process, the role of dendritic cells (Dendritic cells, DC) and T cells at the front end of the immune system is a particularly important link. Dendritic cells are named for the stellate polymorphism, or den...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N15/14
Inventor 李振兴吴烨婷林洪曹立民黄玉浩赵金龙朱文烨
Owner OCEAN UNIV OF CHINA
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