Monoclonal antibody hybridoma cell strain YH4 resisting furaltadone metabolite AMOZ and application thereof

A hybridoma cell line, monoclonal antibody technology, applied in microorganisms, biological testing, biochemical equipment and methods, etc., can solve the problems of high cost, time-consuming, inability to achieve rapid sample detection, etc., and achieve good detection sensitivity and specificity. sexual effect

Inactive Publication Date: 2016-08-31
JIANGNAN UNIV
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Problems solved by technology

[0003] At present, the detection methods of AMOZ are mainly high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS-MS) and other ultraviolet instrument methods, but these methods have the disadvantages of cumbersome operation, time-consuming, and expensive costs, and cannot be realized Rapid detection of a large number of samples

Method used

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  • Monoclonal antibody hybridoma cell strain YH4 resisting furaltadone metabolite AMOZ and application thereof

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Embodiment 1

[0021] Example 1 Preparation of hybridoma cell line YH4

[0022] (1) Synthesis of complete antigen and standard product: Weigh 11.5 mg of o-aldehydobenzoic acid (2-CBA) into a 25 mL round bottom flask, add 1.3 mL of pure water, 60 μL of 1M HCl, add DMF dropwise under magnetic stirring until the Aldehydebenzoic acid is completely dissolved, add furaltadone metabolite AMOZ 30mg, after dissolving, connect the condenser and place it in a water bath at 60°C to reflux overnight, a white precipitate is obtained, filter the reaction solution, put the white solid on the filter paper in Dry in an oven at 37°C to obtain the hapten 2-CP AMOZ. Identification and analysis were carried out by LC / MS.

[0023] The synthesis of the standard product is made by the condensation reaction between the amino group of the nitrofuran metabolite and the aldehyde group of o-nitrobenzaldehyde, and the o-nitrobenzaldehyde is used to replace the o-aldobenzoic acid to obtain the standard product 2- NPAMOZ ...

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Abstract

Belonging to the technical field of food safety immunodetection, the invention discloses a monoclonal antibody hybridoma cell strain YH4 resisting furaltadone metabolite AMOZ and application thereof. The hybridoma cell strain YH4 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.12017. The monoclonal antibody resisting furaltadone metabolite AMOZ is secreted by the hybridoma cell strain YH4. The monoclonal antibody secreted by the cell strain YH4 has good specificity (an IC50 value of 0.2microgram / L) to furaltadone metabolite AMOZ, has good detection sensitivity and affinity, and can be used for specific detection of anti-furaltadone metabolite AMOZ in food safety. The invention also provides a new method for synthesis of furaltadone metabolite AMOZ immunogen, the synthesis steps of hapten are more simplified and effective, thus providing an immunogen synthesis idea and method for future research of people.

Description

technical field [0001] The invention relates to an anti-furaltadone metabolite (AMOZ) monoclonal antibody hybridoma cell line YH4 and its application, belonging to the technical field of food safety immunoassay. Background technique [0002] Furaltadone has a broad antibacterial spectrum and has antibacterial effects on most Gram-positive and negative bacteria, such as Staphylococcus aureus, Escherichia coli, and Streptococcus pyogenes. This product is not easily absorbed in the intestinal tract after oral administration, so it is mainly used for intestinal infection, and can also be used for the treatment of coccidiosis and turkey blackhead. Nitrofuran spectrum antibiotics (furaltadone, furazolidone, nitrofurantoin, nitrofurazone) mainly appear in animal foods. They have harmful effects on human health and are carcinogenic. Many countries including the United States, Canada and the European Union have banned the use of food. These countries have a ban on all imported food...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/64G01N33/577
CPCC07K16/44
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
Owner JIANGNAN UNIV
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