A rapid and simple fluorescence enzyme-linked immunosorbent assay for tetrabromobisphenol A in soil samples based on a bifunctional fusion protein

Ecotoxicol Environ Saf. 2020 Jan 30:188:109904. doi: 10.1016/j.ecoenv.2019.109904. Epub 2019 Nov 6.

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.

Keywords: Alkaline phosphatase; Fluorescent immunoassay; Nanobody; Soil detection; Tetrabromobisphenol A.

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*

Substances

  • Flame Retardants
  • Polybrominated Biphenyls
  • Recombinant Fusion Proteins
  • Single-Domain Antibodies
  • Soil Pollutants
  • Alkaline Phosphatase
  • tetrabromobisphenol A