Single-chain antibody against fenitrothion and preparation method thereof
A technology for single-chain antibodies and fenitrothion, which is applied in the field of detection antibodies, can solve the problems of no preparation of anti-fenitrothion single-chain antibodies, and achieve enhanced library capacity and molecular diversity, improved efficiency, and high specificity sexual effect
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[0029] (1) Preparation of fenitrothion antigen
[0030] Dissolve 5 g of fenitrothion in 40 ml of ether, pour it into a separatory funnel, and use 30 ml of cold 1% Na 2 CO 3 The solution was washed 3 times. Add 40ml of acetic acid-hydrochloric acid solution (volume ratio: 9:1) to the upper solution, and add 8g of zinc powder several times. The yellow reaction mixture was heated and stirred under reflux for 50 min; cooled, filtered, and the precipitate was washed with carbon tetrachloride, the filtrate was combined, extracted twice with carbon tetrachloride, and the filtrate was extracted with 40 ml of double distilled water. After standing for stratification, the upper layer was removed, and the lower layer was anhydrous Na 2 SO 4 Remove water. The next day, after filtration, the brown oily amino-fenitrothion was obtained. Dissolve 500 mg of amino-fenitrothion in 50 ml of distilled water containing 10 mmol of hydrochloric acid, cool, and slowly add cold 0.1 mol / L NaNO 2 ...
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