一种烯啶虫胺新型半抗原的合成与免疫应用  

Synthesis and Immune Application of a New Type of Nitenpyram Hapten

作　　者：赵方方[1] 罗金辉[1] 谢德芳[1] 吕岱竹[1] 

机构地区：[1]中国热带农业科学院分析测试中心海南省热带果蔬产品质量安全重点实验室,海南海口571101

出　　处：《热带作物学报》2016年第7期1394-1398,共5页Chinese Journal of Tropical Crops

基　　金：中国热带农业科学院分析测试中心主任基金项目(No.2013-01);蔬菜产品未知危害因子识别与已知危害因子安全性评估(No.GJFP2015001)

摘　　要：为建立烯啶虫胺(nitenpyram,NIT)残留免疫分析方法,通过合成烯啶虫胺新型半抗原并进行紫外和质谱等技术的分析确证,分别将半抗原与牛血清蛋白(BSA)和卵清蛋白(OVA)偶联成完全的免疫抗原和包被抗原。免疫小鼠后测定抗血清的效价,随着免疫次数的增加先逐渐升高后再逐渐降低,最高达到1∶16 000。选取抗血清效价最高的小鼠,用间接竞争ELISA法测定其灵敏度,其半抑制浓度(IC50)较低,为1.68μg/m L,竞争效果明显。将建立的检测方法与液相色谱方法进行比较,虽线性范围和灵敏度较液相色谱方法稍低,但回收率均≥80%,无明显差异,具有明显的识别效应,为烯啶虫胺高灵敏的单克隆抗体制备及残留免疫分析奠定基础。A new type of nitenpyram hapten was analyzed and confirmed after being synthesized by UV spectrum and mass spetrum in order to establish a method for the analysis of the residues of nitenpyram（NIT）. And the NIT hapten was then coupled with BSA and OVA to prepare immunogen（NIT-BSA） and coating antigen（NITOVA）. The titer of the antiserum was determined by immune mice, and the titer increased and then reduced gradually with the increase of the number of times. The maximum reached 1 ∶ 16 000. The mice with the highest titer were selected, and the sensitivity was determined by indirect competitive ELISA method. The IC50 was 1.68μg/m L, the semi inhibition concentration was low and the competition effect was obvious. Compared with the method of liquid chromatography, the linearity range and sensitivity are slightly lower than that of liquid chromatography. The two recovery rates were more than 80% without obvious difference which showed obvious recognition effect. This study laid a foundation for the preparation of monoclonal antibodies that has the high sensitivity and the immunoassay.

关 键 词：烯啶虫胺 新型半抗原 合成 免疫效果 

分 类 号：TQ450.2[化学工程—农药化工]