Cyclomycin family antibiotic enzyme-linked immunoassay reagent kit
A technology of enzyme-linked immunoassay and detection kit, which is applied in the field of enzyme immunoassay, and can solve the problems of lack of sensitivity and specificity and high pretreatment requirements of microbial inhibition method
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Embodiment 1
[0037] 1 Preparation of coated antigen
[0038] Weigh 1 g of tetracycline hydrochloride and dissolve it in 50 mL of water, add 2.7 g of chlorosuccinimide, react for 30 minutes, filter, and wash the crude product with water. The crude product was treated with 150 mL of water and diethyl ether (volume ratio 1:1), the water phase was discarded, the diethyl ether phase was evaporated to dryness, washed with water, and filtered to obtain the pure product (about 5.6 g), which was modified tetracycline.
[0039]Dissolve 20mg of modified tetracycline and 6mg of succinic anhydride in 2mL of dioxane, react at 37°C for 2 hours, and place at 4°C overnight. This is solution ①. Take another 35mg of ovalbumin (OVA) dissolved in 2mL of water, and then add 1mL of dioxane, this is the ② solution. Then take 125 mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and dissolve it in 2 mL of water. h, slowly drop the reaction solution into ② solution, stir for 4 hours, then a...
Embodiment 2
[0055] Antigen: use oxytetracycline hydrochloride standard substance instead, antibody: use oxytetracycline antibody solution instead. The ELISA plate was coated with oxytetracycline-coated antigen, and the configuration of the remaining reagent bottles was the same as in Example 1, which was the oxytetracycline ELISA kit.
Embodiment 3
[0057] Antigen: use aureomycin hydrochloride standard substance instead, antibody: use aureomycin antibody solution instead. The ELISA plate was coated with aureomycin-coated antigen, and the configuration of the rest of the reagent bottles was the same as in Example 1, which was the aureomycin enzyme-linked immunoassay kit.
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