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An anti-mabuterol monoclonal antibody hybridoma cell line gz03 and its application

A hybridoma cell line and cloned antibody technology, applied in the field of food safety immunology detection, can solve the problems of complicated operation, high cost, time-consuming and the like, and achieve the effects of good sensitivity, good detection sensitivity and specificity

Active Publication Date: 2020-01-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for detection of bromobuterol, clenbuterol and mabuterol are mainly gas chromatography (GC), liquid chromatography (HPLC), spectrophotometry, molecular imprinting, capillary electrophoresis, gas chromatography, Electrochemical immunosensors and other methods, but these methods have disadvantages such as cumbersome operation, time-consuming, and expensive costs, and cannot achieve rapid detection of a large number of samples

Method used

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  • An anti-mabuterol monoclonal antibody hybridoma cell line gz03 and its application
  • An anti-mabuterol monoclonal antibody hybridoma cell line gz03 and its application

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Experimental program
Comparison scheme
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Embodiment 1

[0021] Embodiment 1: Preparation of hybridoma cell line GZ03

[0022] 1. Synthesis of complete antigen

[0023] Weigh 7.81 mg of mabuterol into a small glass bottle, add 68 μL of 1 M HCl solution, then add 1 mL of ultrapure water until completely dissolved, pre-cool at 4°C for 30 min; add 13 μL of 30% HCl solution to the solution NaNO 2 solution, and react in an ice-water bath for 1 h to obtain an activation solution; dilute 10 mg of KLH protein with CB buffer solution; slowly add the activation solution to the carrier protein KLH dropwise, and adjust the pH to 9.0 while adding the activation solution until the color of the reaction solution is Turn dark yellow to stop adding the activation solution; after reacting in an ice bath at 4°C for 4 hours, the complete antigen and uncoupled small molecules were separated by dialysis, and identified by ultraviolet absorption scanning method.

[0024] 2. Animal immunity

[0025] Healthy BALB / C mice aged 6-8 weeks were selected for i...

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Abstract

The invention discloses an anti-mabuterol monoclonal antibody hybridoma cell strain GZ03 and application thereof, and belongs to the field of food security immunological detection. The anti-mabuterol monoclonal antibody hybridoma cell strain GZ03 is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.13091. A cell strain produced monoclonal antibody has good specificity and detection sensitivity on mabuterol, and can be used for residue detection of mabuterol in food, wherein the IC50 value is 0.41 ng / mL.

Description

technical field [0001] The invention relates to an anti-mabuterol monoclonal antibody hybridoma cell line GZ03 and applications thereof, which belong to the field of food safety immunology detection and relate to the hybridoma cell line GZ03 and the anti-mabuterol monoclonal antibody produced therefrom. Background technique [0002] β-adrenoreceptor stimulants are a class of feed additives that can promote the growth of lean meat, including clenbuterol hydrochloride, brobuterol, mabuterol, salbutamol, ractopamine, dopamine hydrochloride, terbutaline sulfate, 15 drugs including phenylethanolamine A, bambuterol, zilpaterol hydrochloride, clorprenaline hydrochloride, sibuterol, arformoterol tartrate, and formoterol fumarate. Among them, clenbuterol hydrochloride was first used as a feed additive and has the widest application range. The chemical structure of Mabuterol (Mab) is very similar to that of Clenbuterol (cl), and it is very likely to be an alternative to Clenbuterol. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577
CPCC07K16/44G01N33/94
Inventor 匡华刘睿胥传来徐丽广马伟刘丽强吴晓玲宋珊珊胡拥明
Owner JIANGNAN UNIV