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Ciprofloxacin hapten, artificial antigen and antibody and preparation method and application thereof

A technology of ciprofloxacin and artificial antigen, applied in the field of food safety, can solve problems such as the use of arms that have not yet been seen, and achieve the effect of high sensitivity

Inactive Publication Date: 2011-02-02
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports of researchers using an arm between the target ciprofloxacin and the carrier protein

Method used

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  • Ciprofloxacin hapten, artificial antigen and antibody and preparation method and application thereof
  • Ciprofloxacin hapten, artificial antigen and antibody and preparation method and application thereof
  • Ciprofloxacin hapten, artificial antigen and antibody and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Hapten Synthesis

[0043]Weigh 38.6mg (about 0.1mmoL) of ciprofloxacin, 11.5mg (about 0.1mmoL) of NHS and 20.3mg (about 0.1mmoL) of DDC into a round-bottomed flask, and dissolve the mixture with a small amount of DMF. Stir overnight in the dark. Centrifuge the reactant at 10,000 r / min for 10 min, and take the supernatant, which is called liquid A. Take 10.3mg (0.1mmoL) of GABA and dissolve it in 1mL of distilled water, namely solution B. Slowly add liquid B to liquid A, stir overnight at room temperature, centrifuge at 10000r / min for 10min, take the supernatant, and wash with saturated NaHCO 3 Adjust to alkaline (pH value 8-8.5), there is precipitation, filter the precipitation, and collect the supernatant. Use dilute hydrochloric acid (1mol / L) to adjust the supernatant to acidic (pH 5-6), collect the precipitate, which is the ciprofloxacin-aminobutyric acid hapten.

Embodiment 2

[0044] Example 2: Hapten Synthesis

[0045] Weigh 38.6mg (about 0.1mmoL) of ciprofloxacin, 11.5mg (about 0.1mmoL) of NHS and 20.3mg (about 0.1mmoL) of DDC into a round-bottomed flask, and dissolve the mixture with a small amount of DMF. Stir overnight in the dark. Centrifuge the reactant at 10,000 r / min for 10 min, and take the supernatant, which is called liquid A. Take 10.3mg (0.1mmoL) of GABA and dissolve it in 1mL of distilled water, namely solution B. Slowly add liquid B to liquid A, stir overnight at room temperature, centrifuge at 10000r / min for 10min, take the supernatant, and wash with saturated NaHCO 3 The pH value was adjusted to 8.2, and a precipitate was precipitated. The precipitate was filtered and the supernatant was collected. Use 1 mol / L dilute hydrochloric acid to adjust the pH of the supernatant to 5.5, and collect the precipitate, which is the ciprofloxacin-GABA hapten.

Embodiment 3

[0046] Example 3: Hapten Synthesis

[0047] Weigh 38.6mg (about 0.1mmoL) of ciprofloxacin, 11.5mg (about 0.1mmoL) of NHS and 20.3mg (about 0.1mmoL) of DDC into a round-bottomed flask, and dissolve the mixture with a small amount of DMF. Stir overnight in the dark. Centrifuge the reactant at 10,000 r / min for 10 min, and take the supernatant, which is called liquid A. Take 0.1mmoL of aminocaproic acid and dissolve it in 1mL of distilled water, namely solution B. Slowly add liquid B to liquid A, stir overnight at room temperature, centrifuge at 10000r / min for 10min, take the supernatant, and wash with saturated NaHCO 3 The pH value was adjusted to 8.5, and a precipitate was precipitated. The precipitate was filtered and the supernatant was collected. Adjust the pH of the supernatant to 5.0 with 1 mol / L dilute hydrochloric acid, and collect the precipitate, which is the ciprofloxacin-GABA hapten.

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Abstract

The invention provides Ciprofloxacin hapten, artificial antigen and antibody and a preparation method and application thereof. Ciprofloxacin is taken as a raw material, and is reacted with aminobutyric or aminocaproic to generate the hapten containing 4 to 6 chiral carbon atoms; the hapten is coupled with protein to prepare the artificial antigen by an active ester method; and the artificial antigen is used to immunize a mouse, and the monoclonal antibody with high specificity on the Ciprofloxacin is prepared by adopting cell fusion technology. The monoclonal antibody is adopted to establish immunological detection for quickly detecting the Ciprofloxacin on site, and has realistic significance for quickly detecting the Ciprofloxacin in livestock, poultry and aquatic products.

Description

technical field [0001] The invention belongs to the technical field of food safety, and in particular relates to a preparation method and application of ciprofloxacin hapten, artificial antigen and antibody. Background technique [0002] Ciprofloxacin (Ciprofloxacin, CPF), its molecular formula is C 17 h 18 o 3 N 3 F, molecular weight is 385.82, is white or off-white crystalline powder, bitter in the mouth, and its structural formula is as shown in formula (I): [0003] [0004] Ciprofloxacin is one of the typical representatives of the third-generation quinolones (FQs), which has the characteristics of broad antibacterial spectrum, low toxicity and side effects, high blood drug concentration, long half-life, various preparation forms, and high drug efficacy and price ratio. Both Gram-negative and positive bacteria have strong antibacterial effects, and have no cross-resistance with penicillins, cephalosporins and amino drugs, and have special effects on the treatment...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D215/56G01N33/577C07K14/77C07K14/765C07K16/44
Inventor 孙远明张挺雷红涛沈玉栋杨金易王弘
Owner SOUTH CHINA AGRI UNIV
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