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Method for sensitive detection of freshwater fish main allergen parvalbumin

A technology for the main allergen and parvalbumin, applied in the special preparation process and detection field of freshwater fish parvalbumin, can solve the problems of unfavorable detection kit development and low yield of parvalbumin, and achieve good separation and repeatability. Good, high yield and high purity results

Active Publication Date: 2012-10-03
JIMEI UNIV
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Problems solved by technology

However, in the past, the research on parvalbumin was mainly based on its properties. The yield of purified parvalbumin was low, or the natural parvalbumin could not be obtained by heating, which was not conducive to the development of later detection kits.

Method used

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  • Method for sensitive detection of freshwater fish main allergen parvalbumin
  • Method for sensitive detection of freshwater fish main allergen parvalbumin
  • Method for sensitive detection of freshwater fish main allergen parvalbumin

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specific Embodiment approach

[0028]1): Preparation of freshwater fish silver carp parvalbumin standard:

[0029] (1) Preparation of parvalbumin crude extract: 20g silver carp white meat was mashed in 4 times the volume of 20mmol / L Tris-HCl (pH 7.5) buffer, centrifuged at 8000g for 30min to obtain parvalbumin crude extract thing. Silver carp parvalbumin crude extract SDS-PAGE analysis results see figure 2 shown.

[0030] (2) Ammonium sulfate salting out: 86 mL crude parvalbumin extract was subjected to 60% ammonium sulfate salting out, that is, 31.05 g of ammonium sulfate was added to the crude extract, stirred for 60 min, left standing for 2 h, and centrifuged at 15000 g for 30 min. Take 95 mL of the supernatant, add 26.51 g of ammonium sulfate, stir for 60 min, let stand for 2 h, and centrifuge at 15000 g for 30 min. The precipitate was dissolved in 10 mL of 20 mmol / L Tris-HCl (pH 7.5), and dialyzed in this buffer for 16 h.

[0031] (3) Ion-exchange column chromatography: load the dialyzed sample on...

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Abstract

The invention discloses a method for sensitive detection of freshwater fish main allergen parvalbumin, which is characterized in that the method comprises the following steps of: 1) preparation of a freshwater fish silver carp parvalbumin standard substance; 2) preparation of an anti-silver carp parvalbumin monoclonal antibody; 3) biotin labeling of the anti-silver carp parvalbumin monoclonal antibody; 4) establishment of a fish parvalbumin competitive ELISA detection method; 5) detection of fish parvalbumin in a sample to be detected. The invention realizes the detection of trace fish parvalbumin in food.

Description

technical field [0001] The invention relates to a method capable of sensitively detecting parvalbumin, the main allergen of freshwater fish, in particular to a special preparation process and detection method of parvalbumin, the main allergen of freshwater fish. Background technique [0002] Food allergy has become an important health problem worldwide. Although various foods may cause allergic reactions in the human body, the currently recognized "main allergenic foods" refer to the eight categories of foods that can cause serious adverse reactions in the body, including milk, eggs, peanuts, tree nuts, crustaceans, fish, etc. beans, wheat and soybeans. Although modern biological and medical related technologies are changing with each passing day, the most effective prevention method for food allergies is still to strictly avoid the intake of allergens. Therefore, it is especially important for allergy sufferers to know the exact composition of food, especially potential a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531G01N33/532
Inventor 蔡秋凤曹敏杰刘光明苏文金
Owner JIMEI UNIV
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