Polymyxin sulfate B monoclonal antibody hybridoma cell strain and application thereof

A technology of polymyxin sulfate and hybridoma cell lines, applied in the direction of polymyxin, antibacterial immunoglobulin, biochemical equipment and methods, etc., can solve the problems of adding colistin, etc., and achieve good detection sensitivity Effect

Inactive Publication Date: 2020-03-27
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because colistin has a broad antibacterial spectrum and significan

Method used

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  • Polymyxin sulfate B monoclonal antibody hybridoma cell strain and application thereof
  • Polymyxin sulfate B monoclonal antibody hybridoma cell strain and application thereof

Examples

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Embodiment 1

[0024] Example 1 Preparation of hybridoma cell line OVA17

[0025] (1) Hapten: The chemical structure of polymyxin B sulfate contains an active group (-NH 2 ) Can be used directly as a hapten.

[0026] (2) Preparation of complete antigen: Weigh 14 mg of PMB, add 1.5 mL of 0.1M PBS to dissolve, add 0.5 mL of tetrahydrofuran solution containing 2.8 mg of GMBS to it after fully dissolving, and stir at 30°C for 30 min. The tetrahydrofuran was removed by nitrogen blowing, and then washed with 3 mL of ether-dichloromethane (V / V=2 / 1) for 3 times to remove excess GMBS, and finally PMB-GMBS solution, namely A solution. Take 10 mg BSA, dissolve it in 0.2 mL 0.1M PBS, and add 50 µL of the 0.5M hydroxylamine solution, stirred at 25°C for 10 min, and then add 3 mL 0.1M PBS (containing 3M urea) to dilute to obtain solution B. Add liquid B to liquid A immediately, and stir vigorously at 25°C for 30 min. The mixed solution was dialyzed with 0.01M PBS solution to remove unreacted small molecule h...

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Abstract

The invention discloses a polymyxin sulfate B monoclonal antibody hybridoma cell strain and application thereof, and belongs to the field of food safety immunodetection. The polymyxin sulfate B monoclonal antibody hybridoma cell strain OVA17 is obtained through screening and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.18516. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 value is 13.13 ng/mL) for polymyxin B sulfate, can realize detection of polymyxin B sulfate residue in feed and livestock and poultry food, provides a raw material for immunodetection of polymyxin B sulfate residue in food. Practical application value is achieved.

Description

Technical field [0001] The invention relates to a polymyxin B sulfate monoclonal antibody hybridoma cell strain and its application, and belongs to the field of food safety immune detection. Background technique [0002] Polymyxin B Sulfate (Polymyxin B) English name: Polymyxin B Sulfate (PMB), is a group of polypeptide antibiotics produced by Bacillus polymyxa. PMB is a white crystalline powder, easily soluble in water, has hygroscopicity, and is stable in acidic solutions. The neutral solution does not affect the potency when placed at room temperature for a week, and it is unstable in alkaline solutions. The antibacterial spectrum and clinical application of Polymyxin B are similar to those of Polymyxin E. It is resistant to Gram-negative bacilli such as Escherichia coli, Pseudomonas aeruginosa, Para Escherichia coli, Klebsiella pneumoniae, Acidophilus, Bacillus pertussis and Shigella has inhibitory or bactericidal effects. Clinically, it is mainly used for infections caused...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/12C07K7/62G01N33/68C12R1/91
CPCC07K7/62C07K16/1278G01N33/68G01N2410/00
Inventor 胥传来李月匡华徐丽广马伟刘丽强吴晓玲宋珊珊胡拥明
Owner JIANGNAN UNIV
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