Ofloxacin couple and its preparing method and use
A technology of ofloxacin and conjugates, applied in the field of antibiotic drug immunodetection, can solve the problems of high price, short shelf life, inability to meet detection needs, etc., and achieves the effect of making up for a long time and saving inspection time.
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Embodiment 1
[0039] (1) Preparation of liquid A: Weigh 10.00 mg ofloxacin, 12.75 mg hydroxysuccinimide, and 26.55 mg EDC, dissolve them in 3ml dimethylformamide, and react to form the active intermediate of ofloxacin and EDC. body, spare;
[0040](2) Preparation of cBSA: Dissolve 18.03mg of ethylenediamine in 20ml of phosphate buffer solution with a pH of 7.40 and a concentration of 0.01M at 0-4°C, adjust the pH to 7.40 with concentrated hydrochloric acid; weigh 1000.00 mg BSA (molecular weight: 68,000) and 57.51 mg EDC, then added to ethylenediamine solution, stirred and reacted at 20°C for 2 hours; The dialyzed solution was stirred and dialyzed for 70 hours, and then dialyzed with distilled water for 24 hours, and the dialysate was replaced every 6 hours; the dialyzed solution was centrifuged at 13,000 rpm at 0-4°C for 15 minutes, and the supernatant was taken; Clear liquid, obtain white powder solid cBSA, standby;
[0041] (3) Preparation of solution B: dissolve cBSA in a phosphate bu...
Embodiment 2
[0046] (1) Preparation of A: Weigh 10.00 mg ofloxacin, 15.94 mg hydroxysuccinimide, and 53.10 mg EDC, dissolve them in 4.6 ml dimethylformamide, and react to form the active intermediate of ofloxacin and EDC body, spare;
[0047] (2) Preparation of cBSA: Dissolve 18.03mg of ethylenediamine in 20ml of phosphate buffer solution with a pH of 7.40 and a concentration of 0.01M at 0-4°C, adjust the pH to 7.40 with concentrated hydrochloric acid; weigh 1000.00 mg BSA (molecular weight: 68,000) and 57.51 mg EDC, then added to ethylenediamine solution, stirred and reacted at 25°C for 4 hours; The dialyzed solution was stirred and dialyzed for 72 hours, and then dialyzed with distilled water for 30 hours, and the dialysate was replaced every 6 hours; the dialyzed solution was centrifuged at 13,000 rpm at 0-4°C for 15 minutes, and the supernatant was taken; freeze-dried Clear liquid, obtain white powder solid cBSA, standby;
[0048] (3) Preparation of solution B: dissolve cBSA in a pho...
Embodiment 3
[0053] (1) Preparation of liquid A: Weigh 10.00 mg ofloxacin, 25.50 mg hydroxysuccinimide, and 79.65 mg EDC, dissolve them in 10 ml dimethylformamide, and react to form the active intermediate of ofloxacin and EDC body, spare;
[0054] (2) Preparation of cBSA: Dissolve 18.52mg of ethylenediamine in 20ml of phosphate buffer solution with a pH of 7.56 and a concentration of 0.02M at 0-4°C, adjust the pH to 7.56 with concentrated hydrochloric acid; weigh 1000.00 mg BSA (molecular weight: 67,000) and 57.51 mg EDC, then added to ethylenediamine solution, stirred and reacted at 22°C for 3 hours; The dialyzed solution was stirred and dialyzed for 80 hours, and then dialyzed with distilled water for 20 hours, and the dialysate was replaced every 6 hours; the dialyzed solution was centrifuged at 13,000 rpm at 0-4°C for 15 minutes, and the supernatant was taken; freeze-dried Clear liquid, obtain white powder solid cBSA, standby;
[0055] (3) Preparation of solution B: dissolve cBSA in...
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