A method for separating egg white allergens

A separation method and egg white technology, which are applied in the preparation methods of peptides, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of inability to separate four allergens

Active Publication Date: 2020-12-18
ZUNYI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the separation and purification methods reported so far can only separate one allergen at a time, and cannot simultaneously separate the four main allergens in egg white

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  • A method for separating egg white allergens
  • A method for separating egg white allergens

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Embodiment Construction

[0015] The present invention will be further described below in conjunction with the examples, but the present invention is not limited to the following examples, it can be foreseen that those skilled in the art may produce various changes in the implementation under the condition of combining the prior art.

[0016] A method for separating egg white allergens, the steps comprising:

[0017] 1. Preparation of egg white treatment solution: shell fresh eggs and separate the egg white, stir slowly with a glass rod in an ice bath until there are no lumps, add an equal volume of distilled water and adjust the pH to 6.0 with 1M hydrochloric acid. After stirring for 3 hours at 4°C , 3000g centrifugation for 10 minutes. After the supernatant was collected, the pH was adjusted to 3.8 with 1M hydrochloric acid, stirred at 4°C for 3 hours, and then centrifuged. The supernatant was frozen at -20°C, which was the egg white treatment solution.

[0018] 2. Preparation of chromatographic co...

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Abstract

The invention discloses a method for separating egg white allergens, and belongs to the technical field of protein separation. The method comprises the steps of 1) pH adjustment and centrifugation preparation of egg white treatment liquid; 2) chromatography column preparation; 3) chromatography separation; 4) protein purity detection; 5), protein concentration. The method adopts a cation exchangechromatography method, egg white is treated and then loaded, four proteins are sequentially eluted by adjusting the pH value and the salt concentration of eluent, and it is verified through protein purity detection that after a BCA method is used for detection, the extraction rates of ovalbumin, ovotransferrin, ovomucoid and lysozyme are 60.0%, 52.1%, 29.6% and 90.2% respectively.

Description

technical field [0001] The invention relates to the technical field of protein separation, in particular to a method for separating egg white allergens. Background technique [0002] The allergens in eggs mainly exist in egg whites. Currently, four major allergens are recognized in egg whites, namely ovalbumin (45 kDa), ovomucoid (28 kDa), ovotransferrin (76 kDa) and lysozyme. (14kDa). Allergen standards are convenient to use, however, many standards have been shown to be contaminated with bacterial endotoxins (LPS). LPS can lead to the imbalance of Th1 / Th2 cytokines, so the content of LPS in allergens should be strictly controlled when establishing sensitized animal models or cell experiments. The isolated and purified allergens are free from LPS contamination and are a good choice for sensitization models. [0003] However, the separation and purification methods reported so far can only separate one allergen at a time, and cannot simultaneously separate the four main a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/77C07K14/79C07K14/81C07K1/18C12N9/36
CPCC07K14/77C07K14/79C07K14/8135C12N9/2462C12Y302/01017
Inventor 麻小娟梁汭
Owner ZUNYI MEDICAL UNIVERSITY
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