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Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran

中国农业科学（英文版）2007，Vol.6Issue(9)：1082-1088,7.

Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran

作者信息

1. Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Livzon Group Reagent Factory, Zhuhai 519060, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China;Key Laboratory of Food Quality and Safety, Education Department of Guangdong Province/Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, P.R. China
折叠

摘要

Abstract

To produce high-affinity monoclonal antibodies against pesticide carbofuran, and the develop immunochemical assays for people's health and environmental protection, the hapten 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy) carbonyl]-amino]-butanoic acid (BFNB) of carbofuran was synthesized and Balb/c mice were immunized by the hapten-carrier (BFNB-bovine serum albumin, BFNB-BSA) conjugates. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by the indirect enzyme-linked immunoabsorbent assay (ELISA), based on BFNB-ovoalbumin conjugates (BFNB-OVA). Purified monoclonal antibody (McAb) was obtained from fluids of ascites, deposited by octanoic acid and ammonium sulfate. The affinity and the specificity of McAb were characterized by ELISA or indirect competitive ELISA. A hybridoma cell line (5D3) secreting anti-carbofuran McAb had been established. The titer of culture medium and ascites was up to 1:2.048 × 103 and 1:1.024 × 106, respectively, and the subtype of the McAb was IgG1. The affinity constant of the McAb was about 2.54 × 109 L mol-1, with an IC50 value of 1.18 ng mL-1 and a detection limit of 0.01 ng mL-1. Cross-reactivity studies showed that the McAb was quiet specific for carbofuran, as among the four analogous compounds, they were all hardly recognized (4.59 × 10-4% for 2,3-dihydro-2,2-dimethyl-7-benzofuranol and less than 3.0 × 10-4% for others). The prepared McAb had a very high affinity and specificity,and it could be used to develop ELISA for rapid determination of carbofuran.

关键词

carbofuran, monoclonal antibodies, immunochemical assays

Key words

carbofuran, monoclonal antibodies, immunochemical assays

分类

农业科技

引用本文复制引用

..Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran[J].中国农业科学（英文版）,2007,6(9):1082-1088,7.

基金项目

This study was supported by the Key Project of Natural Science Foundation of Guangdong Province, China (036842), Key Project of Key Technologies R&D Program of Guangzhou City, China (2004Z2-E0031 and 2004Z1-E0061), Natural Science Foundation of Guangdong Province, China (06300421), National HighTech Research and Development Program of China (863 Program) (2006AA10Z447), and Key Technologies R&D Program during the 10th Five-Year Plan Period,China (2002BA518A06-04). （036842）

中国农业科学（英文版）

ISSN：2095-3119

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