An Enzyme-linked Immunosorbent Assay for Detection of Imidacloprid
To measure imidacloprid(IMI) in water,the hapten(imidacloprid) was conjugated to bovine serum albumin(BSA) with the carbodiimide method to form immunogen IMI-BSA;the hapten imidacloprid was conjugated to ovalbumin(OVA) with mixed carbonic anhydride method to form the coating antigens.Balb/c mice were immunized intraperitoneally with as prepare immunogen.Hybridoma cell lines secreting monoclonal antibodies(McAbs) against imidacloprid,which was obtained through routine hybridoma technology and ELISA screening.The subtype of secreting McAbs was IgG1.The titer of antibody was 1 × 107.After optimization of the ELISA conditions,an indirect ELISA procedure for the determination of imidacloprid was established.The limit of detection was(1.76 ± 0.02) μg/L and IC50 was(15.12 ± 1.28) μg/L.The McAbs had no cross-reaction with some analogues of imidacloprid.Intra-assay RSD reached 4.5%,and inter-assay RSD reached 5.1%.The recoveries obtained by standard imidacloprid addition to the different samples as drinking water,underwater and surface water were 102%,97% and 85%,respectively.The produced monoclonal antibodies and the developed ELISA procedure may become a convenient and satisfied analytical tool for monitoring imidacloprid residues in environmental samples.