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Anti-nicarbazin residual marker DNC monoclonal antibody hybridoma cell strain GW and application thereof

A hybridoma cell line, monoclonal antibody technology, applied in microorganisms, biological material analysis, biochemical equipment and methods, etc., can solve problems such as high cost, residue, and inability to achieve rapid sample detection, and achieve good detection sensitivity and specificity. sexual effect

Inactive Publication Date: 2017-05-31
JIANGNAN UNIV +1
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  • Abstract
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Problems solved by technology

However, as an anti-coccidial drug, it will cause different degrees of residues in poultry muscles and tissues, and most of the drugs are excreted with feces
These manures are used for agricultural fertilization, which has adverse effects on the ecological environment and threatens people's lives and health
[0003] At present, the methods for detecting nicarbazine are mainly instrumental methods such as gas chromatography (GC), liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and liquid phase secondary mass spectrometry (LC-MS / MS). However, these methods have the disadvantages of cumbersome operation, time-consuming, and expensive costs, and cannot realize the rapid detection of a large number of samples. Therefore, it is of great significance to establish a fast and simple nicarbazine residue detection method.

Method used

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  • Anti-nicarbazin residual marker DNC monoclonal antibody hybridoma cell strain GW and application thereof

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Embodiment 1

[0021] Example 1: Preparation of hybridoma cell line GW

[0022] 1. Synthesis of complete antigen

[0023] (1) Synthesis of immune complete antigen: Weigh N-succinyl-L-alanyl-L-alanyl-L-alanine 4-nitroaniline (SAN, CAS: 52299-14-6) 4.86 mg (0.01 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) 5.16 mg (0.025 mmol), N-hydroxysuccinimide (NHS ) 3.0 mg (0.025 mmol), these three drugs were dissolved in 0.5 mL N,N-dimethylformamide (DMF), and the pH was adjusted to about 5.0. After 4 hours of activation at room temperature, the activation solution was added dropwise to 0.05 M carbonate buffer solution in which 6 mg BSA was dissolved, and reacted overnight at room temperature to obtain the immune complete antigen (SAN-EDC-BSA) mixture, which was separated by dialysis Conjugate products and unconjugated small molecule haptens, and identify them by UV absorption scanning method;

[0024] (2) Preparation of heterologous coating: weigh 3.63 mg of glutamyl-γ-ρ-...

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Abstract

The invention relates to an anti-nicarbazin residual marker DNC monoclonal antibody hybridoma cell strain GW and application thereof, and belongs to the technical field of food safety immunodetection. The hybridoma cell strain GW is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 12014. The anti-nicarbazin residual marker DNC monoclonal antibody is generated through secretion of the hybridoma cell strain GW. The monoclonal antibody secreted by the cell strain GW has high specificity (the IC50 value is 5 [mu]g / L) on the nicarbazin residual marker DNC, has high detection sensitivity and affinity, and can be applied to specific detection on the nicarbazin residue in food safety.

Description

technical field [0001] The invention relates to an anti-nicarbazine residue marker DNC monoclonal antibody hybridoma cell line GW and application thereof, belonging to the technical field of food safety immune detection. Involved in the monoclonal cell line GW and its production of anti-nicarbazine residue marker (DNC) monoclonal antibody. Background technique [0002] Nicarbazine is a 1:1 complex composed of dinitrodiphenylurea and hydroxydimethylpyrimidine, in which dinitrodiphenylurea (DNC) is regarded as a marker of nicarbazine residue. As a broad-spectrum, high-efficiency, and stable-performance anticoccidial feed drug additive, it has a strong antiprotozoal effect, and also has a certain control effect on coccidiosis. It is mainly used to prevent chicken cecal coccidia (Eimeria tenella) and heap type, giant, poisonous, Eimeria brucei (small intestinal coccidia). According to the test, medication within 48 hours after infection with coccidia can effectively inhibit th...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577
CPCC07K16/44G01N33/577G01N2430/10
Inventor 匡华丁西胥传来徐丽广马伟吴晓玲刘丽强宋珊珊郑乾坤
Owner JIANGNAN UNIV
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