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Method for screening wheat varieties with low celiac sprue toxicity based on T cell epitopes

A celiac disease and epitope technology, applied in the food and biological fields, can solve the problems of high cost, time-consuming and laborious, and low glutenin extraction efficiency, and achieve the effect of simple operation method, simple operation, and clear and intuitive detection results.

Inactive Publication Date: 2014-03-26
NANCHANG UNIV
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Problems solved by technology

The 60% ethanol extraction method is the earliest and more common extraction method, but the extraction efficiency of this method is not high for the glutenin that also has celiac epitopes. It has been reported that the 60% ethanol extraction method can extract glutenin from each gram of wheat. Extract gluten protein 0.01g, the concentration is 0.33mg / mL
Mass spectrometry has the advantages of high sensitivity, less sample consumption, high mass accuracy and reliable results. However, compared with immunological detection methods, mass spectrometry relies on expensive equipment, is expensive, time-consuming and laborious, and is not suitable for on-site applications in the food industry. Rapid, mass detection

Method used

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  • Method for screening wheat varieties with low celiac sprue toxicity based on T cell epitopes
  • Method for screening wheat varieties with low celiac sprue toxicity based on T cell epitopes

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Embodiment 1

[0021] Example 1: Screening of celiac disease-causing T cell epitopes in different wheat varieties.

[0022] The whole screening process includes extraction of wheat gluten protein, SDS-PAGE of gluten protein combined with PageBlue TM Identification of staining, based on immunoblotting of mAbs against T cell epitopes. The specific implementation process includes the following steps.

[0023] 1. Extraction and identification of gluten protein.

[0024] Eight wheat varieties were selected, Nos. 1-8 were Yanshi 16, Neixiang 184, Zhengzhou 941, Yanzhan 893, Xiaoyan 54, Xinhan 1, Zhengnong 16, Yunong 015 and a control variety Bovictus. Gluten protein is extracted after flour.

[0025] Gluten protein was extracted using a 50% isopropanol / DTT two-step method. Select wheat kernels of various varieties and grind them into fine powder with a mortar, pour them into centrifuge tubes, and use them to extract gluten protein. For the first extraction, extract with 50% (v / v) isopropanol,...

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Abstract

A method for screening wheat varieties with low celiac sprue toxicity based on T cell epitopes comprises the following steps: separating and extracting wheat gluten protein in two steps, performing SDS-PAGE electrophoresis combined with PageBlue TM staining method to identify the gluten protein, detecting celiac sprue T cell epitopes caused by the gluten protein through immunoblotting and other steps. According to the invention, T cell epitopes-based screening method is adopted to screen out the wheat varieties with low celiac sprue toxicity, so that safe and reliable low gluten food can be supplied for celiac sprue patients from the origin of raw materials, and the safety risk of the celiac sprue patients is reduced.

Description

technical field [0001] The field of the invention mainly belongs to the fields of food and biotechnology, and relates to a method for screening wheat varieties with low celiac disease toxicity based on T cell epitopes. Background technique [0002] Celiac disease is a chronic intestinal inflammatory disease caused by an intolerance to wheat, rye, and barley gluten in genetically predisposed individuals. Its incidence rate reaches 1-2% in Europe and the United States, and the global trend is on the rise. There is currently no effective treatment for the disease, and a strict gluten-free diet is the only treatment for patients. However, because there are few gluten-free foods on the market and the quality varies, and many foods that do not contain gluten may also be mixed with gluten during processing, which seriously affects the lives of patients with celiac disease. Therefore, it is of great significance to screen wheat varieties with low celiac toxicity. [0003] Gluten ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N1/28
CPCG01N33/505G01N33/5014G01N33/6839G01N2550/00
Inventor 陈红兵朱晓燕吴志华高金燕袁娟丽李欣佟平杨安树
Owner NANCHANG UNIV
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