[1]李盼,张晓帅,邱雨楼,等.有机磷农药广谱特异性单克隆抗体的制备及特性分析[J].江苏农业学报,2017,(06):1415-1421.[doi:doi:10.3969/j.issn.1000-4440.2017.06.031]
　LI Pan,ZHANG Xiao-shuai,QIU Yu-lou,et al.Production and characterization of broad-specific monoclonal antibody for organophosphorus pesticides recognization[J].,2017,(06):1415-1421.[doi:doi:10.3969/j.issn.1000-4440.2017.06.031]
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有机磷农药广谱特异性单克隆抗体的制备及特性分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年06期
页码:
1415-1421
栏目:
加工贮藏·质量安全
出版日期:
2017-12-30

文章信息/Info

Title:
Production and characterization of broad-specific monoclonal antibody for organophosphorus pesticides recognization
作者:
李盼1; 2; 张晓帅2; 3; 邱雨楼2; 闫莉婷2; 杨茜茹2; 3; 刘贤金2; 张存政2; 许扬1
（1.南昌大学食品学院，中德联合研究院，江西南昌330000；2.江苏省农业科学院食品质量安全与检测研究所，江苏南京210000;3.兰州大学药学院，甘肃兰州730000）
Author(s):
LI Pan1; 2; ZHANG Xiao-shuai2; 3; QIU Yu-lou2; YAN Li-ting2; YANG Qian-ru2; 3; LIU Xian-jin2; ZHANG Cun-zheng2; XU Yang1
(1.State Key Laboratory of Food Science and Technology, Sino-Germany Joint Research Institute, Nanchang University, Nanchang 330000, China；2.The Institute of Quality Safety and Nutrition of Agricultural Products, Jiangsu Academy of Agricultural Sciences, Nanjing 210000, China;3.School of Pharmacy, Lanzhou University, Lanzhou 730000, China)
关键词:
有机磷农药; 单克隆抗体; 间接竞争酶联免疫吸附分析（ic-ELISA）
Keywords:
organophosphorus pesticides (Ops); monoclonal antibody; indirect competitive enzyme-linked immunosorbent assay (ic-ELISA)
分类号:
X592
DOI:
doi:10.3969/j.issn.1000-4440.2017.06.031
文献标志码:
A
摘要:
本研究以化合物4-(二乙氧基硫代磷酸酯基)苯甲酸（BPB）为半抗原，与载体蛋白血蓝蛋白（KLH）偶联后免疫Balb/c小鼠。筛选获得1株杂交瘤细胞株，能够分泌广谱识别有机磷农药的单克隆抗体，而后建立间接竞争酶联免疫检测方法，并进行谷物添加方法验证。结果表明，所获得的单克隆抗体对14种有机磷农药具有识别作用，半抑制浓度（IC50）为 0.165～38.274 μg/ml，交叉反应率（CR）为 5.0%～1 134.0%。其中对蝇毒磷、对硫磷、辛硫磷、喹硫磷4种农药具有较高的识别作用，IC50为 0.165～0.873 μg/ml，交叉反应率为 214.0%～1 134.0%。利用建立的间接竞争酶联免疫检测方法对玉米、小麦样品进行添加回收验证，结果表明，该方法对对硫磷、蝇毒磷、辛硫磷、喹硫磷的回收率为 79.2%～127.3%，变异系数均小于11.0%，所建立的方法可用于谷物中多种有机磷农药残留的快速检测。
Abstract:
Chemical 4-(diethoxyphosphorothioyloxy) benzoic acid （BPB） represented the general structure of organophosphorus pesticides (Ops) was synthesized and used as hapten coupled with carrier protein KLH for Balb/c mice immunization. Through hybridoma technology, several cell lines were selected and one cell line was proved to possess the ability of secreting monoclonal antibody with broad specicity against multiple Ops. Thereafter, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the determination of multiple Ops and employed for the fortified grain sample detection and method validation. The results showed that the monoclonal antibody could broadly recognize 14 Ops, and the cross-reactivity (CR) ranged from 5.0% to 1 134.0%, half maximal inhibitory concentration (IC50) ranged from 0.165 μg/ml to 38.274 μg/ml. It was found that the monoclonal antibody had higher affinity for the parathion, coumaphos, phoxim and quinalphos binding, and IC50 ranged from 0.165 μg/ml to 0.873 μg/ml, and CR ranged from 214.0% to 1 134.0%. An ic-ELISA was developed and validated by spiked maize and rice samples. The results showed that the recoveries ranged from 79.2% to 127.3%, and coefficients of variation (CV) were less than 11%. The established method can be used for the determination of multiple Ops in cereal samples.

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备注/Memo

备注/Memo:
收稿日期：2017-04-20 基金项目：江苏省自然科学基金项目（BK20131333）；江苏省农业科技自主创新基金项目[CX(15)1011] 作者简介：李盼（1991-），女，山东泰安人，硕士研究生，主要从事广谱有机磷农药的基因工程抗体研究。（Tel）18021520996；（E-mail）578365878@qq.com 通讯作者：许杨，（Tel）0791-88329479；（E-mail）xuyang1951@163.com
更新日期/Last Update: 2018-01-03