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Toltrazuril-resistant monoclonal antibody hybridoma cell strain K-5 and application thereof

A hybridoma cell line, monoclonal antibody technology, applied in biochemical equipment and methods, material testing products, instruments, etc., can solve the problems of high cost, inability to achieve rapid on-site detection of a large number of samples, and cumbersome operations, and achieve good detection. Effects of sensitivity and specificity

Active Publication Date: 2017-06-30
JIANGNAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for detecting toltrazuril mainly include gas chromatography (GC), liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), tandem mass spectrometry (LC-MS / MS) and other instrumental methods. Its accuracy and sensitivity can meet the needs very well, but these methods have disadvantages such as cumbersome operation, time-consuming, and expensive costs, and cannot realize rapid on-site detection of a large number of samples. Therefore, a fast and simple detection method for toltrazuril is established method is important

Method used

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  • Toltrazuril-resistant monoclonal antibody hybridoma cell strain K-5 and application thereof
  • Toltrazuril-resistant monoclonal antibody hybridoma cell strain K-5 and application thereof

Examples

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Embodiment 1

[0020] Embodiment 1: Preparation of hybridoma cell line K-5

[0021] 1. Synthesis of complete antigen

[0022] Weigh 4.6 mg toltrazuril hapten (TOLCOOH), 4.4 mg 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinyl Dissolve 2.6 mg of amine (NHS) in 0.6 mL of anhydrous N,N-dimethylformamide (the mixture is called solution A), and stir at room temperature for 4 h. Measure 6.0 mg of hemocyanin (KLH), dilute to 3.0 mg / mL with 0.05 M, pH 9.6 carbonate buffer solution (referred to as solution B), and add solution A to B dropwise under the action of magnetic stirring In solution, react overnight at room temperature to obtain the complete antigen (TOL-KLH), separate the complete antigen and uncoupled small molecule hapten by dialysis, and identify by ultraviolet absorption scanning method; preparation of coating original (TOL-OVA) The method is the same as the preparation of immunogen, just replace KLH with chicken ovalbumin (OVA).

[0023] 2. Anim...

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Abstract

The invention discloses a toltrazuril-resistant monoclonal antibody hybridoma cell strain K-5 and application thereof, and belongs to the field of food safety immunodetection. The toltrazuril-resistant monoclonal antibody hybridoma cell strain K-5 is preserved in the China General Microbiological Culture collection center which is called CGMCC for short, and the preservation number is CGMCC No.13095. A monoclonal antibody excreted by the cell strain has good specificity and detecting sensitivity (the IC50 value is 5.0 ng / mL) on toltrazuril, and can be used for detecting residues of toltrazuril in food safety.

Description

technical field [0001] The invention relates to an anti-toltrazuril monoclonal antibody hybridoma cell line K-5 and its application, which belong to the field of food safety immunology detection, and relate to the hybridoma cell line K-5 and the anti-toltrazuril monoclonal antibody produced therewith Antibody. Background technique [0002] Toltrazuril (TOL) is a new type of triazinone broad-spectrum anticoccidial drug, commonly known as toltrazuril soluble powder. It is off-white to light yellow powder, odorless, soluble in ethyl acetate and dichloromethane, slightly soluble in methanol, insoluble in water. It has a remarkable effect on the prevention and treatment of poultry coccidiosis, and it is effective against Eimeria tenella and Eimeria glandularis, Eimeria major, and Eimeria small Ear coccidia have anti-virus effect. It is widely used in animal husbandry. Its coccidial killing mechanism is: by interfering with the division of the coccidia cell nucleus and mitocho...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577
CPCC07K16/44G01N33/94
Inventor 匡华丁西胥传来徐丽广马伟吴晓玲刘丽强宋珊珊胡拥明郑乾坤
Owner JIANGNAN UNIV
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