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Competitive and noncompetitive phage immunoassays for the determination of benzothiostrobin
Authors:Xiude Hua  Liangliang Zhou  Lu Feng  Yuan Ding  Haiyan Shi  Limin Wang  Shirley J Gee  Bruce D Hammock  Minghua Wang
Institution:1. College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China;2. State & Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China;3. Department of Entomology and UCD Cancer Center, University of California, Davis, CA 95616, United States
Abstract:Twenty-three phage-displayed peptides that specifically bind to an anti-benzothiostrobin monoclonal antibody (mAb) in the absence or presence of benzothiostrobin were isolated from a cyclic 8-residue peptide phage library. Competitive and noncompetitive phage enzyme linked immunosorbent assays (ELISAs) for benzothiostrobin were developed by using a clone C3-3 specific to the benzothiostrobin-free mAb and a clone N6-18 specific to the benzothiostrobin immunocomplex, respectively. Under the optimal conditions, the half maximal inhibition concentration (IC50) of the competitive phage ELISA and the concentration of analyte producing 50% saturation of the signal (SC50) of the noncompetitive phage ELISA for benzothiostrobin were 0.94 and 2.27 ng mL−1, respectively. The noncompetitive phage ELISA showed higher selectivity compared to the competitive. Recoveries of the competitive and the noncompetitive phage ELISAs for benzothiostrobin in cucumber, tomato, pear and rice samples were 67.6–119.6% and 70.4–125.0%, respectively. The amounts of benzothiostrobin in the containing incurred residues samples detected by the two types of phage ELISAs were significantly correlated with that detected by high-performance liquid chromatography (HPLC).
Keywords:Benzothiostrobin  Enzyme-linked immunosorbent assay  Phage-displayed peptide  Phage anti-immunocomplex assay  Noncompetitive immunoassay  Peptidomimetics
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