title: Development of immunoassay based on monoclonal antibody reacted with the neonicotinoid insecticides clothianidin  and dinotefuran.
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&#8211;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.
authors.affiliation: Research & Development Division, HORIBA, Ltd., Minami-ku, Kyoto 601-8510, Japan. mikiko.uchigashima@horiba.com
authors.forename: Mikiko
authors.initials: M
authors.lastname: Uchigashima
authors.forename: Eiki
authors.initials: E
authors.lastname: Watanabe
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Ito
authors.forename: Seiji
authors.initials: S
authors.lastname: Iwasa
authors.forename: Shiro
authors.initials: S
authors.lastname: Miyake
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006146:Guanidines / Q000032:analysis* / Q000276:immunology; D007306:Insecticides / Q000032:analysis* / Q000276:immunology; D000073943:Neonicotinoids; D009574:Nitro Compounds / Q000032:analysis* / Q000276:immunology; D013844:Thiazoles / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D006146:Guanidines; D007306:Insecticides; D000073943:Neonicotinoids; D009574:Nitro Compounds; D013844:Thiazoles; C465368:dinotefuran; C480342:clothianidin