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Danofloxacin couple and its preparing method and use

A technology of danofloxacin and conjugates, which is applied in the field of antibiotic drug immunodetection, can solve the problems of short shelf life, high price, inability to meet detection needs, etc., and achieve the effect of saving inspection time.

Inactive Publication Date: 2006-09-06
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is no report on the synthesis of immunogen and antibody preparation of dafloxacin in the world, and most of the domestic ELISA kits for detecting drug residues of dafloxacin are purchased abroad, with short shelf life and high price. It is far from meeting the needs of detection, so it is necessary to study the synthesis of immunogen and antibody preparation of dafloxacin

Method used

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  • Danofloxacin couple and its preparing method and use
  • Danofloxacin couple and its preparing method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Preparation of liquid A: Weigh 10.00 mg of danofloxacin, 10.13 mg of hydroxysuccinimide, and 21.09 mg of EDC, dissolve them in 3ml of dimethylformamide, and react to form the active intermediate of danofloxacin 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 the ethylenediamine solution, stirred and reacted at 20°C for 2 hours; Phosphate buffer 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; Dry the supernatant to obtain cBSA as a white powder solid, for subsequent use;

[0041] (3) Prepara...

Embodiment 2

[0046] (1) Preparation of A: Weigh 10.00 mg of danofloxacin, 12.66 mg of hydroxysuccinimide, and 42.17 mg of EDC, dissolve them in 4.6 ml of dimethylformamide, and react to form the active intermediate of danofloxacin 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: dis...

Embodiment 3

[0053] (1) Preparation of solution A: Weigh 10.00 mg of danofloxacin, 20.26 mg of hydroxysuccinimide, and 63.26 mg of EDC, dissolve them in 10 ml of dimethylformamide, and react to form the active intermediate of danofloxacin 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 solutio...

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Abstract

The invention discloses a danofloxacin coupling compound with general formula (I), which comprises coupling compounds of danofloxacin hapten and carrier substance of bovine serum albumin which can produce immunogen or egg albumin. Thereinto, n stands for molecular number of danofloxacin combined with a bovine serum albumin molecule, the said n is an integer between one and twenty, and BSA stands for bovine serum albumin with 6.6KDa-6.9KDa of molecular weight. The invention also discloses a process for preparing the said coupling compound, which consists of joining danofloxacin and carrier substance which can produce immunogen to obtain the said coupling compound which induces immune system of animals to produce antibody. By immuning white rabbits from New Zealand, the danofloxacin coupling compound of this invention has prepared antiserum with 1:256,000 of potency, of which the lowest check limit is 0.01ppb. The invention is characterized in that it is simple, rapid, specific, exact and so on, which provides a foundation for preparing enzyme immunoassay agent box of danofloxacin.

Description

technical field [0001] The invention relates to a conjugate of quinolone antibiotics and a preparation method and application thereof, in particular to a conjugate of danofloxacin and a preparation method and application thereof. The invention belongs to the field of antibiotic immunoassay. Background technique [0002] The following titles involved in the present invention apply to the entire specification and claims: [0003] BSA: bovine serum albumin (Bovine Serum Albumin), Sigma company product [0004] PBS: Phosphate Buffered Saline (0.01M, pH=7.40) [0005] Sephadex-675: dextran gel, product of Sigma [0006] cBSA: bovine serum albumin modified with ethylenediamine [0007] Dialysis membrane: product of United Carbide [0008] Dafloxacin: China Veterinary Drug Control Institute [0009] EDC: Ethyl[3-(dimethylamino)propyl]carbodiimide, referred to as EDC, a product of Sigma [0010] Danofloxacin belongs to the quinolone antibiotics. Because of its broad antibact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07D471/08C07K1/00C07K16/18G01N33/53
Inventor 郗日沫卢圣欣张玉兰
Owner SHANDONG UNIV
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