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Preparation method of shrimp tropomyosin allergen primary standard substance

A tropomyosin and allergen technology, applied in peptide preparation methods, chemical instruments and methods, biological tests, etc., can solve the problems of irreproducible results and waste.

Inactive Publication Date: 2014-05-21
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no consensus on the threshold of triggering allergic reactions, and there are great individual differences, but there is a consensus that trace amounts of allergens can cause allergic symptoms in most allergic patients
In order to effectively control shrimp allergens, sensitive and reliable detection methods must be established. Although many units and researchers have established different detection methods based on their own research, the current research on allergens is basically in a state of individualism. The results of different research institutions cannot be replicated, resulting in huge waste

Method used

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  • Preparation method of shrimp tropomyosin allergen primary standard substance

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Effect test

Embodiment 1

[0018] Embodiment 1: The preparation of shrimp tropomyosin allergen reference substance, the steps are as follows:

[0019] (1) extraction

[0020] Shrimp meat was crushed and dissolved in 0.01mol / L, pH7.4, phosphate buffer containing 0.01 mg / mL enzyme inhibitor, the mass of shrimp meat: the volume ratio of phosphate buffer was 1:20, at 4°C Under the condition of magnetic stirring for 10 h, and centrifuged at 5000 r / min for 20 min, the supernatant was collected to obtain the extract of shrimp protein.

[0021] (2) Thermal precipitation

[0022] Heat the shrimp protein extract to 70°C and keep it for 20 minutes, centrifuge at 5000r / min for 20 minutes at 4°C, collect the supernatant, remove heat-labile impurities, and obtain heat-tolerant shrimp class extract, and freeze-dried to obtain heat-resistant shrimp protein powder.

[0023] (3) Saturated ammonium sulfate fractional precipitation

[0024] According to the ammonium sulfate concentration distribution table, prepare an ...

Embodiment 2

[0027] Embodiment 2: SDS-PAGE identification of fish allergen protein

[0028] Shrimp protein extract obtained in (1), (2), (3), (4) steps in Example 1, heat-resistant shrimp protein powder, saturated ammonium sulfate fractionation precipitation components, affinity chromatography The components of each peak were identified by SDS-PAGE. Electrophoresis conditions are: 12% separating gel, 5% stacking gel, constant voltage 120V. figure 1 The standard substance prepared in the method has a single band and a relative molecular weight of 36kDa, which is consistent with the literature report.

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Abstract

The invention discloses a preparation method of shrimp tropomyosin allergen primary standard substance, and belongs to the technical field of biology. The invention relates to a method used for high-efficient stable extraction, separation and purification of main allergen protein tropomyosin from shrimp meat. According to the method, a plurality of technologies, such as immersing extraction, thermoprecipitation, and affinity chromatography are combined for purification of the tropomyosin, the tropomyosin with high allergen performance is extracted from shrimp meat, and original activity is maintained. The tropomyosin obtained via the method is helpful for researches on shrimp allergy, and can be used for immunization technology to detect shrimp allergen in foods.

Description

technical field [0001] The invention discloses a preparation method of a shrimp tropomyosin allergen reference substance, which belongs to the field of biotechnology. Background technique [0002] With the globalization of the world economy and the internationalization of food trade, food allergy has increasingly become a food safety issue of concern. As one of the eight major food allergens, the allergens of crustacean seafood such as shrimp and crab have become the focus of attention. At present, there is no consensus on the threshold of triggering allergic reactions, and there are great individual differences, but there is a consensus that trace allergens can cause allergic symptoms in most allergic patients. In order to effectively control shrimp allergens, sensitive and reliable detection methods must be established. Although many units and researchers have established different detection methods based on their own research, the current research on allergens is basical...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/36C07K1/30C07K1/22G01N33/68
CPCC07K14/43509
Inventor 王国英陈官芝
Owner QINGDAO UNIV
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