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Food allergen dynamic digestion model and in-vitro simulation evaluation method

An in vitro simulation and food allergy technology, applied in biological testing, material inspection products, measuring devices, etc., can solve the cumbersome and complicated process, cannot truly and effectively evaluate the sensitization of food allergens, and cannot completely replace human mast cell lines and other issues to achieve the effect of reducing the ratio of equipment to space stations

Inactive Publication Date: 2019-12-03
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, the RBL-2H3 cell model is mostly used to detect and evaluate the sensitization of food allergens. The RBL-2H3 cell line from Wistar rats can be used as a substitute for human mast cells to study the allergic response of human mast cells. This cell line needs to be used Mouse serum is sensitized, so it is necessary to obtain sensitized serum through animal experiments before cell experiments, the process is cumbersome and complicated, and it cannot completely replace human mast cell lines
[0007] In summary, the existing food allergen sensitization evaluation methods cannot truly and effectively evaluate the sensitization of food allergens

Method used

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  • Food allergen dynamic digestion model and in-vitro simulation evaluation method

Examples

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Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: In vitro simulated dynamic digestion experiment of allergens

[0048] (1) Feed liquid filling: configure the simulated digestive liquid, acid-base adjustment reagents (HCl, NaHCO3) and bile salt solution required in the gastrointestinal digestion process, and store the digestive liquid in a 4° refrigerator before use to protect enzyme activity;

[0049] (2) Software opening and preheating: turn on the power and the instrument enters initialization, confirm that the stepping motor pump, pH sensor, temperature sensor, and constant temperature heating plate (electromagnetic heating plate) are connected normally, click the "Settings" button on the main page, and select "Temperature Control" mode, set the temperature of the heating plate, put the sample to be digested and the simulated gastrointestinal reaction chamber on the temperature-controlled heating plate to preheat for 10 minutes.

[0050] (3) pH Calibration: Click the "Settings" button on the main page to...

Embodiment 2

[0055] Embodiment 2: SDS-polyacrylamide gel experiment

[0056] Simulated gastric digestion products (0, 1, 2, 5, 10, 20, 30, 60, 90) and simulated duodenal intestinal digestion products (0, 1, 5, 10, 15, 30, 60, 90, 120), the structural changes and digestion stability of allergen samples during dynamic digestion in vitro were determined by SDS-polyacrylamide gel experiments.

Embodiment 3

[0057] Example 3: Evaluation of allergen digestion product IgG / IgE binding ability in vitro

[0058] The change of IgG / IgE binding capacity of allergen digestion products during dynamic digestion was analyzed by western blot (Westen-blot). The simulated gastric and duodenal digestion products at each time point were combined with rabbit polyclonal antibody (IgG) or serum antibody (IgE) of patients with shrimp allergy after inactivation treatment for experimental analysis.

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Abstract

The invention discloses a food allergen dynamic digestion model and an in-vitro simulation evaluation method. The method mainly comprises the steps of establishing an allergen dynamic digestion modeland an in-vitro simulation evaluation method, simulating a biochemical reaction condition of the stomach and duodenum of a human body through a digestive system simulation device, providing a method for constructing an allergen in-vitro simulated dynamic digestion experiment in the simulated evaluation method, constructing a Caco-2 (human cloned colonic adenocarcinoma cells) small intestine epithelial cell model to simulate small intestine absorption and transport effects, and constructing a KU812 (human peripheral blood basophilic leukemia cells) cell model to evaluate the ability of mast cell degranulation induced by food allergen digestion and transport products. The method of the invention is suitable for evaluating the digestion and sensitization of a purified food allergen or an allergen extracting solution containing a complex food matrix.

Description

technical field [0001] The invention relates to the field of food allergen allergy evaluation, in particular to a food allergen dynamic digestion model and an in vitro simulation evaluation method. Background technique [0002] Food allergy has become a food safety issue of widespread concern at home and abroad. At present, the focus of food allergen research is on the establishment of faster and more efficient detection methods. There are few studies on the safety evaluation of food allergens and the criteria for judging their allergenicity. How to effectively and quickly evaluate food allergies Protoallergenicity has become an urgent problem to be solved in the field of food safety research. To solve this problem, it is necessary to construct a more complete and reasonable evaluation method for allergen sensitization. Existing evaluation methods mainly include bioinformatics analysis, serological screening, in vitro simulated gastric juice digestion, in vitro cell experi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50
CPCG01N33/5005
Inventor 李振兴郭玉蔓罗晨黄玉浩林洪
Owner OCEAN UNIV OF CHINA
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