title: Development of an enzyme-linked immunosorbent assay for thiacloprid in soil and agro-products with phage-displayed peptide.
abstract: A monoclonal antibody (3A5) that can recognize thiacloprid was produced, and a linear 8-residue peptide phage library was constructed. Six phage-displayed peptides were isolated from the linear 8-residue peptide phage library and a cyclic 8-residue peptide phage library. A phage enzyme-linked immunosorbent assay (ELISA) was developed to detect thiacloprid using a phage-displayed peptide. Under the optimal conditions, the half-maximal inhibition concentration (IC50) and the limit of detection (IC10) of the developed phage ELISA were 8.3 and 0.7 μg/L, respectively. Compared with the conventional ELISA, the sensitivity was improved more than 3-fold. The cross-reactivity (CR) was less than 0.08% for the tested structural analogues and was regarded as negligible. The recoveries of thiacloprid ranged from 80.3% to 116.3% in environmental and agricultural samples, which conformed to the requirements for residue detection. The amount of thiacloprid detected by phage ELISA in the samples was significantly correlated with that detected by high-performance liquid chromatography. The current study indicates that isolating phage-displayed peptides from phage display libraries is an alternative method for the development of a sensitive immunoassay and that the developed assay is a potentially useful tool for detecting thiacloprid in environmental and agricultural samples.
authors.affiliation: College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State and Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yin
authors.affiliation: College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State and Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China.
authors.forename: Xiude
authors.initials: X
authors.lastname: Hua
authors.affiliation: College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State and Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Liu
authors.affiliation: College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State and Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Shi
authors.affiliation: Department of Entomology and UCD Cancer Center, University of California, Davis, Davis, CA 95616, USA.
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Gee
authors.affiliation: College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State and Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China. Electronic address: wangmha@njau.edu.cn.
authors.forename: Minghua
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Entomology and UCD Cancer Center, University of California, Davis, Davis, CA 95616, USA.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001937:Brassica / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D007306:Insecticides / Q000032:analysis* / Q000276:immunology; D057230:Limit of Detection; D018551:Solanum lycopersicum / Q000737:chemistry; D008807:Mice, Inbred BALB C; D000073943:Neonicotinoids; D019151:Peptide Library; D010945:Plants, Edible / Q000737:chemistry*; D011725:Pyridines / Q000032:analysis* / Q000276:immunology; D031989:Pyrus / Q000737:chemistry; D012987:Soil / Q000737:chemistry*; D012989:Soil Pollutants / Q000032:analysis* / Q000276:immunology; D013843:Thiazines / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007306:Insecticides; D000073943:Neonicotinoids; D019151:Peptide Library; D011725:Pyridines; D012987:Soil; D012989:Soil Pollutants; D013843:Thiazines; C417209:thiacloprid
keywords: Enzyme-linked immunosorbent assay; Mimotope; Phage peptide library; Phage-displayed peptide; Thiacloprid