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.
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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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