有机磷农药三唑磷单克隆抗体制备及鉴定

doi:10.7506/spkx1002-6630-201101037

食品科学 ›› 2011, Vol. 32 ›› Issue (1 ): 154-157.doi: 10.7506/spkx1002-6630-201101037

有机磷农药三唑磷单克隆抗体制备及鉴定

韦倩妮¹，黄魁英²，雷红涛^{1 ,*} ，樊晓博³，孙远明¹，徐振林¹，王弘¹，沈玉栋¹，徐怀德³

1. 华南农业大学食品学院，广东省食品质量安全重点实验室 2.广州市微生物研究所 3.西北农林科技大学食品科学与工程学院

收稿日期:2010-04-22 出版日期:2011-01-15 发布日期:2010-12-28
通讯作者: 雷红涛 E-mail:hongtao@scau.edu.cn
基金资助:
国家自然科学基金项目(30700663；20877029)；广东省高等学校科技成果转化项目(cgzhzd0808)；广东省科技计划项目(2009B040500002)

Production and Identification of Monoclonal Antibodies against Organophosphate Insceticide Triazophos

WEI Qian-ni1，HUANG Kui-ying2，LEI Hong-tao1,*，FAN Xiao-bo3，SUN Yuan-ming1，
XU Zhen-lin1，WANG Hong1，SHEN Yu-dong1，XU Huai-de3

1. Key Laboratory of Food Quality and Safety of Guangdong Province, College of Food Science, South China Agricultural University,
Guangzhou 510642, China；2. Guangdong Institute of Microbiology, Guangzhou 510663, China；3. College of Food Science,
Northwest A&F University, Yangling 712100, China

Received:2010-04-22 Online:2011-01-15 Published:2010-12-28
Contact: LEI Hong-tao E-mail:hongtao@scau.edu.cn

摘要/Abstract

摘要：

采用活泼酯法和混合酸酐法将三唑磷半抗原(TZPM-Hap)与牛血清蛋白(BSA)和卵血清蛋白(OVA)偶联，分别制备出一种免疫抗原(TZPM-A-BSA)和两种包被抗原(TZPM-A-OVA 和TZPM-M-OVA)。通过免疫小鼠及细胞融合，并对杂交瘤细胞进行经多次筛选和亚克隆得到1 株产三唑磷单克隆抗体阳性细胞株(FC3)，其抗体亚类为IgM。辛酸- 硫酸铵沉淀法纯化后的抗体经间接ELISA 检测，该方法IC50=9.7μg/L，最低检测限(LOD)为1.0μg/L，定量检测线性范围(IC20～IC80)为2.5～100.0μg/L，与其他有机磷农药结构类似物无明显交叉反应，显示出高度的特异性。该抗体可进一步用于免疫分析试剂盒开发。

关键词: 三唑磷, 单克隆抗体, 酶联免疫吸附检测

Abstract:

In this study, artificial antigens for organophosphate insecticide triazophos including one immunogen and two coating antigens were prepared. The hapten (TZPM-Hap) of organophosphate insecticide triazophos was coupled to bovine serum albumin (BSA) by the active ester method to prepare the artificial immunogen TZPM-A-BSA. Coupling of TZPM-Hap to ovalbumin (OVA) by the active ester method or the mixed anhydride method produced the two coating antigens TZPM-A-OVA and TZPM-M-OVA, respectively. Balb/c mice were immunized with the immunogen, and the splenocytes of the immunized mice were fused with Sp2/0 cells to generate monoclonal antibody (McAbs). A hybridoma cell line (FC3) secreting antitriazophos McAbs was obtained, whose subtype was IgM. Compared with TZPM-M-OVA, better sensitivity in the icELISA format using TZPM-A-OVA was found, which demonstrated 9.7μg/L of IC50, 1.0μg/L of LOD, and 2.5－100.0μg/L of work range (IC20－IC80). The McAbs showed no obvious cross-reactivity with other structural-related organophosphate pesticides. The result indicated that one hybridoma cell line with high specificity and sensitivity was obtained, which will be of importance to develop an ELISA kit for triazophos in the future.

Key words: triazophos, monoclonal antibody, enzyme-linked immunosorbnent assay

中图分类号:

韦倩妮，黄魁英，雷红涛,樊晓博，孙远明，徐振林，王弘，沈玉栋，徐怀德. 有机磷农药三唑磷单克隆抗体制备及鉴定[J]. 食品科学, 2011, 32(1 ): 154-157.

WEI Qian-ni1，HUANG Kui-ying2，LEI Hong-tao1,*，FAN Xiao-bo3，SUN Yuan-ming1，. Production and Identification of Monoclonal Antibodies against Organophosphate Insceticide Triazophos[J]. FOOD SCIENCE, 2011, 32(1 ): 154-157.

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有机磷农药三唑磷单克隆抗体制备及鉴定

Production and Identification of Monoclonal Antibodies against Organophosphate Insceticide Triazophos

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