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Tytuł pozycji:

An Ultrasensitive Bioluminescent Enzyme Immunoassay Based on Nanobody/Nanoluciferase Heptamer Fusion for the Detection of Tetrabromobisphenol A in Sediment.

Tytuł :
An Ultrasensitive Bioluminescent Enzyme Immunoassay Based on Nanobody/Nanoluciferase Heptamer Fusion for the Detection of Tetrabromobisphenol A in Sediment.
Autorzy :
Li Z; Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, California 95616, United States.
Wang Y; Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, California 95616, United States.; Department of Pesticides Science, School of Resources and Environment, Anhui Agricultural University, No. 130 Changjiang West Road, Hefei 230036, China.
Vasylieva N; Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, California 95616, United States.
Wan D; Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, California 95616, United States.
Yin Z; Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, California 95616, United States.
Dong J; Shenzhen Forward Pharma Co., Ltd., Shenzhen 518057, China.
Hammock BD; Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, California 95616, United States.
Pokaż więcej
Źródło :
Analytical chemistry [Anal Chem] 2020 Jul 21; Vol. 92 (14), pp. 10083-10090. Date of Electronic Publication: 2020 Jul 07.
Typ publikacji :
Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
Język :
English
Imprint Name(s) :
Original Publication: Washington, American Chemical Society.
MeSH Terms :
Environmental Monitoring*
Luminescent Measurements*
Geologic Sediments/*chemistry
Luciferases/*chemistry
Polybrominated Biphenyls/*analysis
Single-Domain Antibodies/*chemistry
Immunoenzyme Techniques ; Luciferases/metabolism
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Grant Information :
P42 ES004699 United States ES NIEHS NIH HHS; R35 ES030443 United States ES NIEHS NIH HHS
Substance Nomenclature :
0 (Polybrominated Biphenyls)
0 (Single-Domain Antibodies)
EC 1.13.12.- (Luciferases)
FQI02RFC3A (tetrabromobisphenol A)
Entry Date(s) :
Date Created: 20200620 Date Completed: 20210215 Latest Revision: 20210215
Update Code :
20210218
PubMed Central ID :
PMC7780847
DOI :
10.1021/acs.analchem.0c01908
PMID :
32559059
Czasopismo naukowe
Tetrabromobisphenol A (TBBPA) is a flame retardant and has become a widely concerning environmental pollutant. An ultrasensitive nanobody-based immunoassay was developed to monitor the exposure of TBBPA in sediment. First, the anti-TBBPA nanobody was fused with nanoluciferase, and then a one-step bioluminescent enzyme immunoassay (BLEIA) was developed with high sensitivity for TBBPA, with a maximum half inhibition concentration (IC 50 ) at 187 pg/mL. Although approximately 10-fold higher sensitivity can be achieved by this developed BLEIA than by the classical two-step ELISA (IC 50 at 1778 pg/mL), it is still a challenge to detect trace TBBPA in sediment samples reliably due to the relatively high matrix effect. To further improve the performance of this one-step BLEIA, a C4b-binding protein (C4BP) was inserted as a self-assembling linker between the nanobody and nanoluciferase. Therefore, a heptamer fusion containing seven binders and seven tracers was generated. This reagent improved the binding capacity and signal amplification. The one-step heptamer plus BLEIA based on this immune-reagent shows an additional 7-fold improvement of sensitivity, with the IC 50 of 28.9 pg/mL and the limit of detection as low as 2.5 pg/mL. The proposed assay was further applied to determine the trace TBBPA in sediment, and the recovery was within 92-103%. Taking advantage of this heptamer fusion, one-step BLEIA can serve as a powerful tool for fast detection of trace TBBPA in the sediment samples.

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