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Antibody against fluoroquinolones and application thereof

A technology for quinolones and drugs, which is applied in the field of quinolone drug antibodies, can solve the problems of narrow antibacterial spectrum and clinical application range, complicated production process, and long time consumption, and achieve high-throughput rapid detection, simple storage, and high sensitivity.

Active Publication Date: 2011-01-26
北京英太格瑞检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first generation of quinolones, such as nalidixic acid and pyrrolic acid, etc., are not used now because of their narrow antibacterial spectrum and clinical application range
The second generation of quinolones, pipemidic acid is the main domestic application variety, and its antibacterial spectrum has been expanded, but it is only used for urinary tract and intestinal infections, and it is rarely used at present
The preparation of monoclonal antibodies or polyclonal antibodies must be obtained through cell culture. The entire production process is complicated, time-consuming, expensive, and difficult to operate

Method used

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  • Antibody against fluoroquinolones and application thereof
  • Antibody against fluoroquinolones and application thereof
  • Antibody against fluoroquinolones and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, preparation and function test of antibody

[0056] 1. Preparation of quinolone single-chain antibody

[0057] (1) Antibody screening

[0058] Six-month-old male Balb / c mice were taken, and the total RNA of splenocytes was extracted by Trizol one-step method, purified to obtain mRNA, and then reverse-transcribed to obtain cDNA. Using a complete set of primers, the heavy chain variable region (VH) and light chain variable region (VL) genes were respectively amplified using cDNA as a template. Splicing into a single-chain antibody gene (ScFv), then linking the ScFv with the vector pCANTAB5E to obtain pCANTAB5E / ScFv, and transforming Escherichia coli TG1 to obtain a mouse-derived non-immune single-chain antibody library. Phage particles with specific single-chain antibodies against multiple quinolones were screened by ELISA, and their corresponding nucleotide sequences were obtained by sequencing.

[0059] The coding gene sequence of the single-chain antibo...

Embodiment 2

[0089] Embodiment 2, the enzyme-linked immunosorbent assay kit that detects quinolone drug and preparation thereof

[0090] The sample diluent in this experiment: it is the sample diluent in the test kit described in the experiment.

[0091] Plate washing solution in this experiment: it is the washing solution in the test kit described in Experiment 1.

[0092] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0093] 1. ELISA plates coated with ciprofloxacin and carrier protein conjugates;

[0094] 2. Quinolone antibody: the single-chain antibody described in Example 1. The concentration of the antibody working solution is 5.0 ng / mL, and the antibody working solution is obtained by diluting the purified antibody in Example 1 with the sample diluent;

[0095] 3. Standard quinolone: ​​the standard quinolone is ciprofloxacin (CIP), the concentration of the standard solution is 0μg / L, 1μg / L, 3μg / L, 9μg / L, 27μg / L, 81μg / L; Floxacin (CIP) was pur...

Embodiment 3

[0146] Embodiment 3, the test paper that detects quinolone medicine and its preparation and application

[0147] 1. The structure of the test paper

[0148] The test paper is composed of a sample absorbent pad, a colloidal gold pad, a reaction film and a water absorbent pad;

[0149] The sample absorption pad, the colloidal gold pad, the reaction membrane and the water absorption pad are sequentially connected, the end of the sample absorption pad is connected to the beginning of the colloidal gold pad, the end of the colloid gold pad is connected to the beginning of the reaction membrane, and the end of the reaction membrane is connected to the beginning of the reaction membrane. The beginning of the absorbent pad is connected;

[0150] Colloidal gold-labeled quinolone drug antibody is coated on the colloidal gold pad;

[0151] Detection area and quality control area are arranged on described reaction film, and detection area (C line) and quality control area (T line) all a...

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Abstract

The invention discloses an antibody against fluoroquinolones and application thereof. The single-chain antibody consists of a heavy chain variable region, a short peptide for connecting the heavy chain variable region and a light chain variable region, and the light chain variable region connected in turn; the amino acid sequence of the heavy chain variable region is shown as the first position to the 118th position amino acid residues from the N terminal in a sequence 1; and the amino acid sequence of the light chain variable region is shown as the 134th position to 248th position amino acid residues from the N terminal in the sequence 1. A kit and a colloid gold test paper card have the advantages of high sensitivity, high accuracy, high precision, low cost, simple operation, short detection time, suitability for various units, simple storage and long quality guarantee period, can quickly detect massive samples simultaneously and can realize on-site high-flux quick detection. Therefore, the antibody, the kit, the colloid gold test paper and the detection method have important effect of detecting fluoroquinolones.

Description

technical field [0001] The invention relates to a quinolone drug antibody and its application. Background technique [0002] Quinolones (4-quinolones, QNs), also known as pyroxic acids or pyridonic acids, are a class of synthetic antibacterial drugs that mainly act on Gram-negative bacteria. Quinolones are divided into one, two, three, and four generations according to the invention and their antibacterial properties. The first generation of quinolones, specific varieties include nalidixic acid and pyrrolic acid, etc., are not used now because of their narrow antibacterial spectrum and clinical application range. The second generation of quinolones, pipemidic acid is the main domestic application variety, and its antibacterial spectrum has been expanded, but it is only used for urinary tract and intestinal infections, and it is rarely used at present. The third-generation quinolones include a series of drugs such as norfloxacin. The antibacterial spectrum is further expand...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C12N15/13C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10G01N33/566G01N33/577G01N33/531
Inventor 沈建忠吴小平江海洋李杰超王战辉徐飞刘丹程晶王奇昌李娜师伟杨丽丽温凯
Owner 北京英太格瑞检测技术有限公司
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