Carbetapentane citrate hapten, antigen, antibody, detection device and preparation method
A technology of pentoxyverine citrate and a detection device, which is applied in the field of drug safety detection, can solve the problems of high technical requirements for detection personnel, failure to meet rapid detection, and high requirements for detection instruments, and achieve economical speed, easy production, and low cost. cheap effect
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Embodiment 1
[0026] The preparation method of pentoxyverine citrate hapten:
[0027] S1, add 2.0g of α-phenylcyclopentylacetic acid and 30ml of thionyl chloride to a 100ml single-necked bottle in turn, heat at ℃, stir until reflux, reflux for 3-4h, TLC monitors the reaction material α-phenylcyclopentyl Glycolic acid disappeared, lowered to room temperature and stirred, the reaction solution was distilled to dryness under reduced pressure at 50°C, 10ml was added with dichloromethane, stirred until the solution was clear, and α-phenylcyclopentylacetyl chloride was prepared for use;
[0028] S2, add 3.0g of diethylene glycol acetal and 5ml of dichloromethane to another 100ml one-necked bottle successively, stir at room temperature, after mixing evenly, turn to stirring under ice bath, drop the above-mentioned α-phenylcyclopentylacetyl chloride into Wherein, after dropping, turn to room temperature and stir for 1 h, then raise the bath temperature to 40° C. and stir for 5 h, then lower to room...
Embodiment 2
[0031] The preparation method of pentoxyverine citrate immune antigen:
[0032]Take 0.1 mmol of the pentoxyverine citrate hapten prepared in Example 1 and dissolve it in 2 ml of dimethylformamide (DMF), stir and add 0.2 mmol of dicyclohexylcarbodiimide (DCC) and 0.1 mmol of N- Hydroxysuccinimide (NHS), reacted overnight with magnetic stirring at 4°C, centrifuged and took the supernatant to prepare liquid A; weigh 140mg of bovine serum albumin (BSA) and dissolve it in 10ml with a concentration of 0.1mol / L, pH Add 1ml of DMF solution to the PBS solution with a value of 8.0, stir and dissolve to prepare liquid B; under magnetic stirring, drop liquid A into liquid B drop by drop, react at 4°C for 12h, after centrifugation, take the supernatant, and put it in Dialyze with normal saline at 4°C for 3 days, change the dialysate 3 times a day, and prepare the pentoxyverine citrate immune antigen, that is, the pentoxyverine citrate-BSA antigen. The BSA antigen was dispensed into 0.5mL ...
Embodiment 3
[0034] The preparation method of pentovir citrate antigen for detection line:
[0035] Take 0.1mmol of the pentoxyverine citrate hapten prepared in Example 1 and dissolve it in 2ml of DMF, add 0.2mmol of DCC and 0.1mmol of NHS with stirring, react overnight with magnetic stirring at 4°C, centrifuge the supernatant overnight, and prepare Obtain liquid A; weigh 120 mg of chicken ovalbumin (OVA) and dissolve it in 10 ml of PBS solution with a concentration of 0.1 mol / L and a pH value of 8.0, add 1 ml of DMF solution, stir and dissolve to prepare liquid B; under magnetic stirring, dissolve liquid A Drop into solution B drop by drop, react at 4°C for 12h, centrifuge, take the supernatant, dialyze with normal saline at 4°C for 3 days, change the dialysate 3 times a day, and use citrate penetrant to obtain the detection line Verin antigen, that is, pentoxyverine citrate-OVA antigen, the pentoxyverine citrate-OVA antigen is divided into 0.5ml centrifuge tubes at a concentration of 10m...
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