Development of an enzyme-linked immunosorbent assay for thiacloprid in soil and agro-products with phage-displayed peptide.
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.
['Yin|Wei|W|', 'Hua|Xiude|X|', 'Liu|Xiaofeng|X|', 'Shi|Haiyan|H|', 'Gee|Shirley J|SJ|', 'Wang|Minghua|M|', 'Hammock|Bruce D|BD|']
D000818:Animals D000911:Antibodies, Monoclonal D001937:Brassica D004797:Enzyme-Linked Immunosorbent Assay D005260:Female D007306:Insecticides D057230:Limit of Detection D018551:Lycopersicon esculentum D008807:Mice, Inbred BALB C D000073943:Neonicotinoids D019151:Peptide Library D010945:Plants, Edible D011725:Pyridines D031989:Pyrus D012987:Soil D012989:Soil Pollutants D013843:Thiazines
Enzyme-linked immunosorbent assay Thiacloprid Phage-displayed peptide Mimotope Phage peptide library