Lomefloxacin conjugate and its preparing method and use

A technology of lomefloxacin and conjugates, which is applied in the field of immunoassay of antibiotic drugs, can solve the problems of short shelf life, high price, and inability to meet the detection needs, and achieve the effect of saving inspection time

Inactive Publication Date: 2008-09-17
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After retrieval, there is no report on the synthesis of the immunogen of lomefloxacin and the preparation of antibodies in the world, and most of the domestic ELISA kits for detecting drug residues of lomefloxacin are purchased abroad, with short shelf life and high price, far from being available. To meet the detection needs, it is very necessary to study the synthesis of immunogen and antibody preparation of lomefloxacin

Method used

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  • Lomefloxacin conjugate and its preparing method and use
  • Lomefloxacin conjugate and its preparing method and use

Examples

Experimental program
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Effect test

Embodiment 1

[0039] (1) Preparation of liquid A: Weigh 10.00 mg of lomefloxacin, 13.12 mg of hydroxysuccinimide, and 27.31 mg of EDC, dissolve them in 3 ml of dimethylformamide, and react to generate active intermediates of lomefloxacin and EDC. 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 ...

Embodiment 2

[0046] (1) Preparation of A: Weigh 10.00 mg of lomefloxacin, 16.40 mg of hydroxysuccinimide, and 54.63 mg of EDC, dissolve them in 4.6 ml of dimethylformamide, and react to generate an active intermediate of lomefloxacin and EDC, 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 mgBSA (molecular weight 68,000) and 57.51mg EDC, then added to the ethylenediamine solution, stirred and reacted at 25°C for 4 hours; the reaction solution of ethylenediamine and BSA was mixed with the above-mentioned phosphate buffer Stir and dialyze for 72 hours, then dialyze with distilled water for 30 hours, replace the dialysate every 6 hours; centrifuge the dialyzed solution at 13,000 rpm for 15 minutes at 0-4°C, take the supernatant; freeze-dry the supernatant Liquid, obtain white powder solid cBSA, stan...

Embodiment 3

[0053] (1) Preparation of liquid A: Weigh 10.00 mg of lomefloxacin, 26.24 mg of hydroxysuccinimide, and 81.95 mg of EDC, dissolve them in 10 ml of dimethylformamide, and react to generate active intermediates of lomefloxacin and EDC. 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 0.02 M 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: d...

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Abstract

The invention discloses a lomefloxacin coupling compound with general formula (I), which comprises coupling compounds of lomefloxacin hapten and bovine serum albumin of carrier substance which can produce immunogen or egg albumin. Thereinto, n stands for molecular number of lomefloxacin 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 lomefloxacin 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 lomefloxacin coupling compound of this invention has prepared antiserum with 1:6400 of potency, of which the lowest check limit is 1ppb. 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 lomefloxacin.

Description

technical field [0001] The invention relates to a conjugate of quinolone antibiotics and its preparation method and application, in particular to a lomefloxacin conjugate and its preparation method and application. 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 (0.01 M, pH=7.40) [0005] Sephadex-G75: dextran gel, product of Sigma [0006] cBSA: bovine serum albumin modified with ethylenediamine [0007] Dialysis membrane: product of United Carbide [0008] Lomefloxacin: China Veterinary Drug Control Institute [0009] EDC: Ethyl[3-(dimethylamino)propyl]carbodiimide, referred to as EDC, a product of Sigma [0010] Lomefloxacin (lomefloxacin) belongs to the quinolone antibiotics. Because of its broad antib...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C07D401/04C07K1/00C07K16/18G01N33/53
Inventor 郗日沫卢圣欣张玉兰
Owner SHANDONG UNIV
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