A hybridoma cell line dnc secreting anti-triclabendazole monoclonal antibody and its application
A technology of triclobendazole mono-hybridoma cell line, which is applied in the directions of hybrid peptide, analytical material, biological material analysis, etc., can solve the problems of complicated operation, high cost, restrict the popularization of detection method, etc., and achieves simplified synthesis steps, The effect of good detection sensitivity and affinity
Active Publication Date: 2021-10-29
JIANGNAN UNIV
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Problems solved by technology
Instrumental detection methods can perform quantitative analysis and have a low detection limit, but usually require expensive instruments and complicated operations, and the pretreatment and detection time is long, which seriously restricts the promotion of these detection methods
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Embodiment 1
[0028] Embodiment 1: Preparation of anti-triclobendazole monoclonal antibody hybridoma cell line DNC
[0029] 1. Synthesis of hapten: the synthetic route is as follows: figure 1 shown
[0030] First, 5 g of TCD was dissolved in 100 mL of dichloromethane / tetrahydrofuran (v / v, 1:1), 4 g of 3-chloroperbenzoic acid was added, and the resulting mixed solution was stirred at room temperature for 3 hours. Then 100mL 1M Na 2 S 2 o 3 The solution was added to the above solution and the aqueous phase was extracted three times with dichloromethane. The combined organic part was washed with 200 mL of brine, and concentrated under reduced pressure to obtain compound C1, which was precipitated as a yellow solid;
[0031] Dissolve 5 g of compound C1 in 100 mL of DMF, add 4 g of NaHS, and stir at 90° C. overnight. After the reaction, 300 mL of deionized water was added, and the aqueous phase was extracted three times with dichloromethane. The combined organic portion was washed with 20...
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A hybridoma cell line DNC secreting anti-triclobendazole monoclonal antibody and its application belong to the field of food safety immune detection. The hybridoma cell line DNC secreting anti-triclobendazole monoclonal antibody of the present invention has been preserved in the General Microorganism Center CGMCC of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.17395. The monoclonal antibody produced by the secretion of the hybridoma cell line DNC has better affinity and higher sensitivity to triclabendazole, and the 50% inhibitory concentration IC of triclabendazole 50 It is 1.77ng / mL, and can be used to prepare immunoassay kits and colloidal gold test strips for triclabendazole, providing powerful detection methods and means for the detection of triclabendazole in food of animal origin.
Description
technical field [0001] The invention relates to a hybridoma cell line DNC secreting anti-triclobendazole monoclonal antibody and an application thereof, belonging to the field of food safety immune detection. Background technique [0002] Triclobendazole is a new type of benzimidazole anthelmintic, which has a good killing effect on Fasciola hepatica, Fasciola large and Anteroposterior disc fluke at various developmental stages in cattle and sheep, and has low toxicity. Therefore, it is widely used in veterinary clinics to prevent and treat trematodes in ruminant livestock. [0003] There have been a few reports at home and abroad on the detection methods of triclabendazole drug residues in animal tissues, mainly using high performance liquid chromatography-fluorescence detector detection and liquid chromatography-tandem mass spectrometry. Extraction methods are also different, there are liquid-liquid extraction, liquid-solid extraction and so on. Instrumental detection me...
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IPC IPC(8): C12N5/20C07K16/44C07K14/795G01N33/577C12R1/91
CPCC07K14/795C07K16/44C07K19/00C07K2317/92G01N33/577G01N2430/10
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