Journal
JOURNAL OF HAZARDOUS MATERIALS
Volume 424, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jhazmat.2021.127411
Keywords
1-Naphthol; Carbaryl; Nanobody; Ratiometric fluoroimmunoassay; Pesticide exposure
Categories
Funding
- National Key Research and Development of China [2018YFC1602904]
- Science and Technology Foundation of Guangdong Province [2018A050506072]
- Key Project of Guangdong Provincial High School [2019KJDXM002]
- National Institutes of Environmental Health Sciences Superfund Research Program [P42ES04699]
- NIEHS RIVER Award [R35 ES030443-01]
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This study developed a nanobody-alkaline phosphatase fusion protein and a ratiometric fluomimmunoassay (RFIA) for highly sensitive determination of 1-naphthol, showing a 220-fold sensitivity improvement compared to conventional ELISA. A detection system based on silicon nanoparticles and gold nanoclusters was utilized, achieving good experimental results.
The excessive use of carbaryl has resulted in the risk of its exposure. In this study, we isolated six nanobodies (Nbs) from a camelid phage display library against the biomarker of carbaryl, 1-naphthol (1-NAP). Owing to its characteristics of easy genetic modifications, we produced a nanobody-alkaline phosphatase (Nb-CC4-ALP) fusion protein with good stability. A dual-emission system based ratiometric fluomimmunoassay (RFIA) for quick and highly sensitive determination of 1-NAP was developed. Silicon nanoparticles (SiNPs) was used as an internal reference and for aggregation-induced emission enhancement (AIEE) of gold nanoclusters (AuNCs), while AuNCs could be quenched by MnO2 via oxidation. In the presence of ALP, ascorbic acid phosphate (AAP) can be transformed into ascorbic acid (AA), the later can etch MnO2 to recover the fluorescence of the AuNCs. Based on optimal conditions, the proposed assay showed 220-fold sensitivity improvement in comparison with conventional monoclonal antibody-based ELISA. The recovery test of urine samples and the validation by standard HPLC-FLD demonstrated the proposed assay was an ideal tool for screening 1-NAP and provided technical support for the monitoring of carbaryl exposure.
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