Application of Current Hapten in the Production of Broad Specificity Antibodies Against Organophosphorus Pesticides |
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Authors: | LIU Xian-jin YAN Chun-rong LIU Yuan YU Xiang-yang ZHANG Cun-zheng |
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Institution: | Food Safety Research and Inspection Center, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P.R. China |
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Abstract: | Diethylphosphono acetic acid (DPA) was used as a current hapten to generate broad specificity polycolonal antibodies against a group of organophosphorus pesticides. Six New Zealand white rabbits were immunized with immunogens synthesized by the active ester method (AEM) or 1-ethyl-3-(3-dimethylaminopropyl)-carbodimide method (EDC). The titers of antisera reached 25 600 by AEM and 6 400 by EDC, respectively. Polyclonal antibodies raised against DPA were screened and selected for the competitive indirect enzyme-linked immunosorbent assay (CI-ELISA). A CI-ELISA for DPA was developed with a detection limit of 3.536 ng mL-1 and an I50 value of 0.182 ug mL-1. The assay specificity was evaluated by obtaining competitive curves for several structurally related compounds as competitors. The antiserum showed high affinities to chlorpyrifos, diazinon, omethoate, parathion-ethyl and profenofos with I50 of 0.12, 0.15, 0.21, 0.88, 0.97 and 2.5 ug mL-1, respectively. The results indicate that the assay could be a screening tool for quantitation and semi-quantitation determination of the above former five organophosphorus pesticides. |
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Keywords: | organophosphorus pesticides broad specificity antibody enzyme-linked immunosorbent assay (ELISA)LIU Xian-jin Professor Tel: 86-25-84390401 E-mail:jaasliu@jaas ac cn |
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