Title: A rapid and simple fluorescence enzyme-linked immunosorbent assay for tetrabromobisphenol A in soil samples based on a bifunctional fusion protein.
Authors: Fu, Hui-Jun; Wang, Yu; Xiao, Zhi-Li; Wang, Hong; Li, Zhen-Feng; Shen, Yu-Dong; Lei, Hong-Tao; Sun, Yuan-Ming; Xu, Zhen-Lin; Hammock, Bruce
Published In Ecotoxicol Environ Saf, (2020 Jan 30)
Abstract: Tetrabromobisphenol A (TBBPA) is the largest brominated flame retardant which can be released to environment and cause long-term hazard. In this work, we developed a rapid and highly sensitive fluorescence enzyme-linked immunosorbent assay (FELISA) for monitoring of TBBPA in soil samples. TBBPA specific nanobody derived from camelid was fused with alkaline phosphatase to obtain the bi-functional fusion protein, which enable the specific binding of TBBPA and the generation of detection signal simultaneously. The assay showed an IC50 of 0.23 ng g-1, limit detection of 0.05 ng g-1 and linear range from 0.1 to 0.55 ng g-1 for TBBPA in soil samples. Due to the high resistance to organic solvents of the fusion protein, a simple pre-treatment by using 40% dimethyl sulfoxide (DMSO) as extract solvent can eliminate matrix effect and obtain good recoveries (ranging from 93.4% to 112.4%) for spiked soil samples. Good relationship between the results of the proposed FELISA and that of liquid chromatography tandem mass spectrometry (LC-MS/MS) was obtained, which indicated it could be a powerful analytical tool for determination of TBBPA to monitor human and environmental exposure.
PubMed ID: 31704326
MeSH Terms: Alkaline Phosphatase/genetics; Alkaline Phosphatase/metabolism; Animals; Camelids, New World; Environmental Monitoring/methods*; Enzyme-Linked Immunosorbent Assay*; Flame Retardants/analysis*; Limit of Detection; Polybrominated Biphenyls/analysis*; Polybrominated Biphenyls/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Single-Domain Antibodies/genetics; Single-Domain Antibodies/metabolism; Soil Pollutants/analysis*