摘要
该文制备了农药稻瘟灵的单克隆抗体,并建立了稻瘟灵的酶联免疫吸附(ELISA)检测方法。在完全保留稻瘟灵结构的基础上从二硫杂环戊烷结构中衍生不同长度的活性手臂制备了2个半抗原,并分别与载体蛋白偶联合成免疫原与包被原。通过小鼠免疫、细胞融合、淘筛、腹水制备等步骤获得特异性识别稻瘟灵的单克隆抗体mAb-DWL。结果显示,基于mAb-DWL构建的间接竞争ELISA法的半抑制浓度(IC_(50))为55.2 ng/mL,线性范围为4.6~530.2 ng/mL,其与结构类似物的交叉反应可忽略不计。所建立的ELISA方法对蔬菜及粮食等样品的加标回收率为77.2%~116%,可用于实际样品的快速检测。
An enzyme-linked immunosorbent assay(ELISA)method based on the monoclonal anti⁃body was established for determination of pesticide isoprothiolane.Two haptens were prepared by ex⁃tending active arms of different lengths from disulfide heterocyclic pentane structure of isoprothiolane molecule.The two haptens were conjugated with carrier protein to obtain the immunogen and coating antigen.Monoclonal antibody,named mAb-DWL,against isoprothiolane was obtained via the procedures of mouse immunization,cell fusion,eluting sieve and ascites collection.Results showed that the ELISA based on the obtained mAb-DWL exhibited a linear range of 4.6-530.2 ng/mL,with an IC_(50) of 55.2 ng/mL.Meanwhile,the cross-reactivity rates for the mAb-DWL with oth⁃er similar pesticides were lower than 1%.The spiked recoveries for isoprothiolane in the vegetables and grains samples ranged from 77.2%to 116%.The monoclonal antibody prepared by this method could be used for development of rapid method for isoprothiolane determination.
作者
王幸幸
王耀宇
李晓娜
申超群
周凯
张静
WANG Xing-xing;WANG Yao-yu;LI Xiao-na;SHEN Chao-qun;ZHOU Kai;ZHANG Jing(Shenzhen Total-testing Technology Co.,Ltd.,Shenzhen 518038,China;Institute of Jiangxi Oil-tea Camellia,Jiujiang University,Jiujiang 332005,China;Technology Center of Yantai Customs,Yantai 264000,China)
出处
《分析测试学报》
CAS
CSCD
北大核心
2022年第11期1696-1701,共6页
Journal of Instrumental Analysis
基金
江西省教育厅科学技术研究项目(GJJ211804)
深圳市同类检测科技有限公司科研(建设)项目(RD20210001)。
