Six phage-displayed peptides that specifically bind to anti-imidaclothiz monoclonal antibody (mAb) were obtained by biopanning the phage-displayed peptide libraries. A phage enzyme linked immunosorbent assay (P-ELISA) and a phage chemiluminescent enzyme immunoassay (P-CLEIA) for imidaclothiz were developed by using a clone L4-29. The optimal buffers of P-ELISA and P-CLEIA were pH 8, PBS buffer containing 0.14 M NaCl and 2.5% methanol. Under the optimal conditions, the half maximal inhibition concentration (IC50) values, limit of detection (LOD, IC10 values) and linear range (IC10 to IC90) of the P-ELISA were 1.45, 0.55 and 0.55 to 3.82 ng mL-1, while the P-CLEIA were 0.86, 0.13 and 0.13 to 5.84 ng mL-1, respectively. The sensitivities of P-ELISA and P-CLEIA were improved more than 60-fold and 100-fold respectively than the conventional ELISA. The P-CLEIA showed higher sensitivity and wider linear range than P-ELISA. Two analyses had no significant cross-reactivity with the analogues of imidaclothiz except for imidacloprid. Recoveries of the P-ELISA and P-CLEIA for imidaclothiz in paddy water, soil, cabbage, rice, apple, pakchoi, pear and tomato samples were 72.3-101.3% and 73.9-102.6%, respectively. The amounts of imidaclothiz in the containing incurred residues samples detected by the P-ELISA and P-CLEIA were significantly correlated with that detected by high-performance liquid chromatography (HPLC).

Ultra-sensitive enzyme immunoassays for the determination of imidaclothiz using phage-displayed peptide

254th National Meeting (2017)

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