| Production and Characterization of Polyclonal Antibody for the N- Methylcarbamate Insecticide Metolcarb |
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| 作者单位： | ZHANG Qi（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
LI Tie-jun（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
ZHU Xiao-xia（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
XU Li-na（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
LIU Feng-quan（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
HU Bai-shi（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
JIANG Ying-hua（Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China） ;
CAO Bin（Pesticide Research Institute of Jiangsu Province,Nanjing 210036,P.R.China） ; |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | The hapten, 3-{1-(3-(methyl)phenyloxy)-carbonyl]amino}propanoic acid (HOM), mimicking the analyte metolcarb, was synthesized and verified by mass spectrometry (MS) and 1H-nuclear magnetic resonance spectrometry (1H-NMR). Then,HOM was conjugated with the carrier proteins bovine serum albumin (BSA) and ovalbumin (OVA) with stoichiometric amounts of N-hydroxysuccinimide/dicyclohexylcarbodimide (NHS/DCC) using the activated ester method. Polyclonal antibodies were raised against the conjugate of HOM-BSA in rabbits. Antiserum titres were determined by noncompetitive indirect ELISA procedures and the titer of pAb01 reached 1.28 × 106. The cross-reactivities of the structurally related Nmethylcarbamate insecticides were 0.0% except for dimethacarb. These results indicate that the antibody pAb01 with strong affinity and high specificity can be used to develop a sensitive and rapid detection protocol for metolcarb residue.
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| 关 键 词： | N-甲基氨基甲酸酯 杀虫剂 分析方法 多克隆抗体 ELISA |
| 收稿时间： | 26 June 2006 |
Production and Characterization of Polyclonal Antibody for the N-Methylcarbamate Insecticide Metolcarb |
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| Authors: | ZHANG Qi LI Tie-jun ZHU Xiao-xia XU Li-na LIU Feng-quan HU Bai-shi JIANG Ying-hua CAO Bin |
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| Institution: | 1. Key Laboratory of Monitoring and Management of Plant Diseases and Pests,Ministry of Agriculture/Department of Plant Protection,Nanjing Agricultural University,Nanjing 210095,P.R.China
2. Pesticide Research Institute of Jiangsu Province,Nanjing 210036,P.R.China |
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| Abstract: | The hapten, 3-{1-(3-(methyl)phenyloxy)-carbonyl]amino}propanoic acid (HOM), mimicking the analyte metolcarb. was synthesized and verified by mass spectrometry (MS) and 1H-nuclear magnetic resonance spectrometry (1H-NMR). Then, HOM was conjugated with the carrier proteins bovine serum albumin (BSA) and ovalbumin (OVA) with stoichiometric amounts of N-hydroxysuccinimide/dicyclohexylcarbodimide (NHS/DCC) using the activated ester method. Polyclonal antibodies were raised against the conjugate of HOM-BSA in rabbits. Antiserum titres were determined by noncompetitive indirect ELISA procedures and the titer of pAb01 reached 1.28 × 106. The cross-reactivities of the structurally related N-methylcarbamate insecticides were 0.0% except for dimethacarb. These results indicate that the antibody pAb01 with strong affinity and high specificity can be used to develop a sensitive and rapid detection protocol for metolcarb residue. |
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| Keywords: | metolcarb antigen polyclonal antibody antiserum characterization ELISA |
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