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Hybridoma cell line secreting anti-chlorpromazine monoclonal antibody and application thereof

A hybridoma cell line, chlorpromazine single technology, applied in biochemical equipment and methods, material testing products, microorganisms and other directions, can solve the problem of expensive instruments, complicated sample processing, difficult to be widely used in large-scale high-throughput detection, etc. problems, to achieve the effect of simplified synthesis steps, good detection sensitivity and affinity

Inactive Publication Date: 2019-05-03
JIANGNAN UNIV +1
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Problems solved by technology

[0003] At present, the conventional methods for detecting chlorpromazine residues include spectrophotometer, fluorescence spectrometry, liquid chromatography, liquid chromatography-mass spectrometry, and electrochemical methods. These methods or instruments are expensive, or sample processing is complicated, and relatively Difficult to be widely used in large-scale high-throughput detection

Method used

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  • Hybridoma cell line secreting anti-chlorpromazine monoclonal antibody and application thereof
  • Hybridoma cell line secreting anti-chlorpromazine monoclonal antibody and application thereof
  • Hybridoma cell line secreting anti-chlorpromazine monoclonal antibody and application thereof

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Embodiment 1

[0020] Embodiment 1: Preparation of anti-chlorpromazine monoclonal antibody hybridoma cell line D

[0021] 1. Synthesis of hapten: Acepromazine maleate (APZ, 45 mg, 0.01 mmol) and carboxymethoxamine hemihydrochloride (CMO, 25 mg, 0.02 mmol) were dissolved in deionized water (5 mL) , and sodium acetate was added dropwise while the rotor was stirring to adjust the pH to 8-9, then the mixture was refluxed at 40°C for 10 h, the pH was adjusted to 2-3 with 0.1 M hydrochloric acid, and then 15 mL of ethyl acetate was added. The ethyl acetate layer was collected and dried with a rotary evaporator to obtain chlorpromazine hapten (CPZ-hapen).

[0022] 2. Synthesis of complete antigen: take 2.2mg of chlorpromazine hapten and dissolve it in 2mL of dimethylformamide, add 4.5mg of EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide salt acid) and 3.6mg NHS (N-hydroxysuccinimide), stirred at room temperature, and activated for 4 hours; another 10mg KLH (keyhole limpet hemocyanin) was dissol...

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Abstract

The invention relates to a hybridoma cell line secreting an anti-chlorpromazine monoclonal antibody and an application thereof, which belong to the field of food safety immunoassay. The hybridoma cellline D secreting the anti-chlorpromazine monoclonal antibody prepared by the invention is preserved in the General Microbiology Center of the China Microbial Culture Collection Management Committee,and the preservation number is CGMCC No. 17304. The hybridoma cell line D has a good affinity and high sensitivity to chlorpromazine, the IC50 of the 50% inhibitory concentration of chlorpromazine is17.2 [mu]g / L, the hybridoma cell line can be used for the preparation of a chlorpromazine immunoassay kit and a colloidal gold test strip, which provides a powerful detection method and means for thedetection of chlorpromazine in animal-derived foods.

Description

technical field [0001] The invention relates to a hybridoma cell strain secreting anti-chlorpromazine monoclonal antibody and application thereof, which belongs to the field of food safety immune detection. Background technique [0002] Chlorpromazine (CPZ), also known as Dongmianling, belongs to phenothiazine drugs and is a central dopamine receptor blocker with sedative, antipsychotic, hypothermia and basal metabolism effects. In veterinary medicine, this drug is mainly used for sedation and antiemetic. Adding chlorpromazine to the diet can make animals indirectly fatten. Some unscrupulous traders often use large doses of chlorpromazine during animal transportation. This is because chlorpromazine can reduce the maintenance needs of animals, reduce weight loss and mortality rate. Large doses of chlorpromazine used before slaughtering can lead to a large amount of chlorpromazine remaining in animal products. The accumulated residue of chlorpromazine in the animal body will...

Claims

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Application Information

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IPC IPC(8): C07K16/44C12N5/20G01N33/53
Inventor 胥传来王忠兴匡华徐丽广刘丽强马伟吴晓玲宋珊珊胡拥明
Owner JIANGNAN UNIV
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