期刊
ANALYTICAL BIOCHEMISTRY
卷 572, 期 -, 页码 9-15出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2019.02.022
关键词
One-step immunoassay; Carbaryl; Chicken single-chain variable fragment; Alkaline phosphatase; Pesticide monitoring
资金
- Key Project of Intergovernmental International Scientific and Technological Innovation Cooperation [2016YFE0108900]
- National Key Research and Development Program of China [2016YFD0800606]
- National Institute of Environmental Health Sciences Superfund Research Program [P42ES04699]
Immunoassays provide a high-throughput method for monitoring pesticides in foods and the environment. Due to easy generation and capable of being manipulated, chicken single-chain variable fragment (scFv) is attractive in the development of immunoassays for pesticides. Two scFvs (X1 and X2) against the insecticide carbaryl were generated from a chicken immunized with hapten Cl conjugated to keyhole limpet hemocyanin and fused with alkaline phosphatase (AP) to develop a rapid one-step enzyme-linked immunosorbent assay for this pesticide. X2-AP showed higher binding affinity to carbaryl than X1-AP. The X2-AP-based ELISA had a half-maximum signal inhibition concentration of 15 ng mL(-1) and a limit of detection of 1.6 ng mL(-1). This assay showed negligible cross-reactivity with other carbamate pesticides (< 0.1%) and low cross-reactivity with 1-naphthol (5%). The average recoveries of carbaryl spiked in soil, apple and pear samples by the one-step assay ranged from 90% to 114% and agreed well with those of high-performance liquid chromatography. The chicken scFv-based assay showed promise as a high-throughput screening tool for carbaryl in environmental and food matrices.
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