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Development of Immunoassay Based on Monoclonal Antibody Reacted with the Neonicotinoid Insecticides Clothianidin and Dinotefuran
Research & Development Division, HORIBA, Ltd., Minami-ku, Kyoto 601-8510, Japan
National Institute for Agro-Environmental Sciences, Tsukuba, Ibaraki 305-8604, Japan
Department of Materials Science, Toyohashi University of Technology, Toyohashi, Aichi 441-8580, Japan
Advanced Scientific Technology & Management Research Institute of Kyoto, Shimogyo-ku, Kyoto 600-8813, Japan
* Author to whom correspondence should be addressed.
Received: 12 October 2012; in revised form: 12 November 2012 / Accepted: 13 November 2012 / Published: 15 November 2012
(This article belongs to the Special Issue Bioassays
Abstract: Enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody (MoAb) was developed for the neonicotinoid insecticide clothianidin. A new clothianidin hapten (3-[5-(3-methyl-2-nitroguanidinomethyl)-1,3-thiazol-2-ylthio] propionic acid) was synthesized and conjugated to keyhole limpet hemocyanin, and was used for monoclonal antibody preparation. The resulting MoAb CTN-16A3-13 was characterized by a direct competitive ELISA (dc-ELISA). The 50% of inhibition concentration value with clothianidin was 4.4 ng/mL, and the working range was 1.5–15 ng/mL. The antibody showed high cross-reactivity (64%) to dinotefuran among the structurally related neonicotinoid insecticides. The recovery examinations of clothianidin for cucumber, tomato and apple showed highly agreement with the spiked concentrations; the recovery rate was between 104% and 124% and the coefficient of variation value was between 1.8% and 15%. Although the recovery rate of the dc-ELISA was slightly higher than that of HPLC analysis, the difference was small enough to accept the dc-ELISA as a useful method for residue analysis of clothianidin in garden crops.
Keywords: enzyme-linked immunosorbent assay; ELISA; hapten
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Uchigashima, M.; Watanabe, E.; Ito, S.; Iwasa, S.; Miyake, S. Development of Immunoassay Based on Monoclonal Antibody Reacted with the Neonicotinoid Insecticides Clothianidin and Dinotefuran. Sensors 2012, 12, 15858-15872.
Uchigashima M, Watanabe E, Ito S, Iwasa S, Miyake S. Development of Immunoassay Based on Monoclonal Antibody Reacted with the Neonicotinoid Insecticides Clothianidin and Dinotefuran. Sensors. 2012; 12(11):15858-15872.
Uchigashima, Mikiko; Watanabe, Eiki; Ito, Shigekazu; Iwasa, Seiji; Miyake, Shiro. 2012. "Development of Immunoassay Based on Monoclonal Antibody Reacted with the Neonicotinoid Insecticides Clothianidin and Dinotefuran." Sensors 12, no. 11: 15858-15872.