Preparation of bionic molecular imprinting electrochemical sensor and based on click chemistry and detection of food allergen

A technology of molecular imprinting and click chemistry, applied in the direction of material electrochemical variables, etc., can solve problems such as expensive, complex antibody preparation process, maximum detection limit limit application and development, etc.

Inactive Publication Date: 2013-02-27
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most widely used method is the skin test, which can provide rapid but non-quantitative results for allergy diagnosis, and in vitro diagnosis is undoubtedly convenient and reliable for both doctors and patients, but due to ELISA test kits and Test strips have a complicated antibody preparation process and are relatively expensive; although PCR technology has development prospects, the denaturation of DNA during food processing and the maximum detection limit of this method limit its current application and development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Detection of Peanut Allergens

[0032] (1) Weigh 30g of peanuts, use a tissue crusher to crush them into powder, immerse in n-hexane to degrease at a weight-to-volume ratio of g / mL 1:10, leaching overnight at 4°C, refrigerate and centrifuge, collect the upper layer of protein infusion, 10mL of protein Slowly add the immersion solution to saturated ammonium sulfate with a mass concentration of 30%, let it stand overnight at 4°C, centrifuge and precipitate, then slowly add the supernatant to a saturated ammonium sulfate with a mass concentration of 50%, let it stand for 30 minutes, and centrifuge to dissolve 30%- Dissolve the precipitated protein fraction at 50% saturation in PBS, add sulfuric acid to the isoelectric point of pH 5.2, dissolve the precipitate in PBS after centrifugation, put it into a dialysis bag and dialyze against distilled water for 4 hours, change the dialysate once during the period, and then transfer to Continue dialysis into PBS to obtai...

Embodiment 2

[0042] Example 2: Detection of egg allergens

[0043] (1) Separate the egg white from the egg yolk, weigh 30g of egg white, immerse in n-hexane to degrease according to the weight-to-volume ratio g / mL of 1:10, stir the egg white evenly, extract overnight at 4°C, refrigerate and centrifuge, and collect the upper protein Infusion solution, 10mL protein infusion solution was slowly added to saturated ammonium sulfate with a mass concentration of 30%, stood overnight at 4°C, centrifuged to precipitate, and then slowly added to the supernatant to saturated ammonium sulfate with a mass concentration of 50%, stood for 30min, and centrifuged , Dissolve the precipitated protein components with 30%-50% saturation in PBS, add sulfuric acid to the isoelectric point of pH 5.2, dissolve the precipitate in PBS after centrifugation, put it into a dialysis bag and dialyze against distilled water for 4 hours, and change the dialysis during the period solution once, and then transfer to PBS to c...

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PUM

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Abstract

The invention discloses preparation of a bionic molecular imprinting electrochemical sensor and based on click chemistry and a detection method of food allergen. The method includes selecting nitrine alkyl sulfhydryl fixed with the surface of an electrode to prepare a self-assembly single layer film of a nitrine end group, utilizing click chemistry reaction to conduct end group alkenyl functionalization on the self-assembly single layer film on the nitrine end group, selecting a function single body capable of reacting with the allergen to combine molecularly imprinted polymers (MIPs) of an allergen artificial antibody, utilizing the existing method to prepare a graphene material, mixing the graphene material to prepare the artificial antibody, evenly mixing template molecules of the food allergen, the function single body, the graphene, crosslinked agent, pore-foaming agent, initiator and organic solvent according to certain substance amount to prepare the bionic MIPs mingled with the graphene, connecting a prepared MIP modification work electrode to an electrochemical working station and detecting the food allergen in food extraction liquid.

Description

technical field [0001] The present invention relates to the technical field of food allergen detection, more specifically to the preparation of an electrochemical sensor capable of detecting food allergens, and the present invention also relates to a method for determining trace food allergens in a sample using the bionic molecular imprinting technology . Background technique [0002] Food allergens have become an emerging public food safety issue, especially in developed countries, surveys show that more than 1% of adults worldwide are allergic to food, and the incidence of infants and children has reached 5%- 10%. Generally speaking, food allergens are proteins or glycoproteins belonging to different families, and a small amount of food allergen proteins can cause severe allergic reactions. In recent years, the International Codex Alimentarius Commission and various developed countries have issued various food allergen labeling regulations, standards and corresponding im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26
Inventor 葛慎光颜梅于京华葛磊黄加栋张彦楚成超王盼盼刘芳刘伟艳孙国强
Owner UNIV OF JINAN
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