中国农业科学 ›› 2005, Vol. 38 ›› Issue (03): 511-515 .

• 植物保护 • 上一篇    下一篇

吡虫啉人工抗原的合成与鉴定

朱国念,桂文君,程敬丽,郑尊涛   

  1. 浙江大学农药与环境毒理研究所
  • 收稿日期:2003-12-12 修回日期:1900-01-01 出版日期:2005-03-10 发布日期:2005-03-10
  • 通讯作者: 朱国念

Synthesis and Identification of Artificial Antigen for Imidacloprid

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  1. 浙江大学农药与环境毒理研究所
  • Received:2003-12-12 Revised:1900-01-01 Online:2005-03-10 Published:2005-03-10

摘要: 以吡虫啉原药与β-巯基丙酸为起始原料,在碱性条件下反应,合成了半抗原{1-[6-(2-羧基乙硫基)-3-吡啶基甲基]-N-硝基亚咪唑烷-2-叉胺}。利用该半抗原与牛血清蛋白(BSA)和卵清白蛋白(OVA)偶联得到了人工抗原,经免疫动物后,获得了高效价的特异性多克隆抗体,抗血清效价为2.56×104。经酶联免疫反应(ELISA)测定,吡虫啉对抗体的抑制中浓度(IC50)为 20.7 ?g·L-1,最低检测限(IC20 )为1.1 ?g·L-1,检测范围在1~1 000 ?g·L-1内线性关系较好,吡虫啉结构类似物交叉反应率均小于1.4%。

关键词: 吡虫啉, 人工抗原, 抗体

Abstract: This study reported that a hapten of imidacloprid, 1-[(6-Carboxylethylthio-3-pyridinyl)methyl]-N-nitro- imidazolidinimine was synthesized by using technical grade imidacloprid reacted withβ-Mercaptopropionic acid in the hot alkalic solution. The hapten was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the artificial antigens, respectively. The antibody with high titer (2.56×104) was produced after immuning to rabbits. Results showed that the antibody hadhigh affinity to imidacloprid tested by enzyme linked immunosorbent assay (ELISA), which IC50 and IC20 were 20.7 ?g·L-1 and 1.1 ?g·L-1 ; and the crosss-reactibities to the related compounds wereless than 1.4%.

Key words: Imidacloprid, Artificial antigen, Antibody