Development of an enzyme-linked immunosorbent assay for thiacloprid in soil and agro-products with phage-displayed peptide

作者：Yin, Wei; Hua, Xiude; Liu, Xiaofeng; Shi, Haiyan; Gee, Shirley J.; Wang, Minghua^*; Hammock, Bruce D.

来源：Analytical Biochemistry, 2015, 481: 27-32.

DOI：10.1016/j.ab.2015.04.015

• Thiacloprid
• Enzyme-linked immunosorbent assay
• Phage-displayed peptide
• Phage peptide library
• Mimotope

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 mu g/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.

• 出版日期2015-7-15
• 单位南京农业大学