A camelid VHH-based fluorescence polarization immunoassay for the detection of tetrabromobisphenol A in water

Wang, Kai; Liu, Zhiping; Pei, Jian; Liu, Jianfeng; Eremin, Sergei A.; Li, Qing X.; Li, Ji; Xu, Ting

doi:10.1039/c6ay01603k

Cited by 7 publications

(3 citation statements)

References 38 publications

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“…Despite these challenges, consumers would welcome a fast test to detect contamination by bisphenol A. Compared to instrument-based methods, immunoassays have advantages such as simplicity, specificity, low chemicals consumption, and high sensitivity, making them a desirable choice for rapid detection [8,9]. Various sensor platforms, particularly electrochemical ones, have shown successful analytical performance using aptamers.…”

Section: Introductionmentioning

confidence: 99%

BPA Endocrine Disruptor Detection at the Cutting Edge: FPIA and ELISA Immunoassays

Raysyan,

Zwigart,

Eremin

et al. 2023

Biosensors

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BPA is a chemical commonly used in the production of polymer-based materials that can have detrimental effects on the thyroid gland and impact human reproductive health. Various expensive methods, such as liquid and gas chromatography, have been suggested for detecting BPA. The fluorescence polarization immunoassay (FPIA) is an inexpensive and efficient homogeneous mix-and-read method that allows for high-throughput screening. FPIA offers high specificity and sensitivity and can be carried out in a single phase within a timeframe of 20–30 min. In this study, new tracer molecules were designed that linked the fluorescein fluorophore with and without a spacer to the bisphenol A moiety. To assess the influence of the C6 spacer on the sensitivity of an assay based on the respective antibody, hapten–protein conjugates were synthesized and assessed for performance in an ELISA setup, and this resulted in a highly sensitive assay with a detection limit of 0.05 g/L. The lowest limit of detection was reached by employing the spacer derivate in the FPIA and was 1.0 μg/L, working range from 2 to 155 μg/L. The validation of the methods was conducted using actual samples compared to LC–MS/MS, which served as the reference method. The FPIA and ELISA both demonstrated satisfactory concordance.

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Section: Introductionmentioning

confidence: 99%

BPA Endocrine Disruptor Detection at the Cutting Edge: FPIA and ELISA Immunoassays

Raysyan,

Zwigart,

Eremin

et al. 2023

Biosensors

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show abstract

“…Fluorescence polarization immunoassay (FPIA) is a homogeneous method without multiple separations or washing steps, which allows for a high-throughput, rapid, and quantitative analysis of the chemical contaminants within a few minutes. In the past few years, FPIA has been developed for monitoring veterinary drugs [ 16 , 17 ], environmental pollutants [ 18 , 19 , 20 ], pesticides [ 21 , 22 , 23 ], and toxins [ 24 , 25 , 26 ]. For more than a decade, our group has reported many FPIA methods to detect hazardous compounds, such as aflatoxins, zearalenone, sulfamethazine, orbifloxacin, and amantadine.…”

Section: Introductionmentioning

confidence: 99%

Dual-Wavelength Fluorescence Polarization Immunoassay for Simultaneous Detection of Sulfonamides and Antibacterial Synergists in Milk

Duan

Zhang

et al. 2022

Biosensors

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Combinations of sulfonamides (SAs) and antibacterial synergists (ASGs) are frequently used for treating infectious diseases and promoting growth for animals, which cause potential hazards to food safety and human health. To realize the simultaneous detection of SAs and ASGs in food, a homogeneous and high-throughput screening dual-wavelength fluorescence polarization immunoassay (DWFPIA) was developed. In this study, three SAs tracers and three ASGs tracers were synthesized by fluoresceins with different linkers and paired with their corresponding monoclonal antibodies (mAbs), respectively. To achieve a high sensitivity and broad specificity, the combination of tracers SADMPM-HDF with the longest linker paring mAb 10E6 for SAs and tracer HaptenA-DSCA paring mAb 9C9 for ASGs were chosen for the development of DWFPIA, achieving surprising IC50 values for 23 SAs below 100 μg L−1 and 5 ASGs below 50 μg L−1. The accuracy of DWFPIA was applied in real milk samples by typical sulfamethazine (SMZ) and trimethoprim (TMP), with recoveries of 81.7–97.2% and 78.6–103.6%, and coefficient of variations (CVs) below 18.9%, which could be completed within 15 min, including sample pretreatment. We firstly developed a simultaneous screening DWFPIA, covering all of the SAs and ASGs used in clinic and providing a great application potential in food safety analysis.

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“…VHHs are used in various fields including diagnostics and therapeutics due to their features such as small size, high stability, , cost-effective production using phage display method, − and high accessibility to antigens. While VHHs have been used in competitive FPIAs for small molecules, − noncompetitive FPIAs for large biomolecules have not been investigated. As an extension of FPIA to the quantification of large molecules, applying a 15 kDa VHH to a 150 kDa protein FPIA is expected to give a 60% Δ P , which is a huge improvement on the fundamental FPIA drawback.…”

Section: Introductionmentioning

confidence: 99%

Noncompetitive Fluorescence Polarization Immunoassay for Protein Determination

et al. 2020

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Fluorescent polarization immunoassay (FPIA) is a single-step immunoassay method that is applicable to point-of-care testing; however, its applicability to large biomolecules has been restricted because ordinary FPIA is a competitive assay. Here, we report a noncompetitive FPIA using the variable domain from the heavy chain of a camelid antibody (VHH antibody). FPIA with VHH was successfully used to quantitate rabbit immunoglobulin G (IgG) and demonstrated a wider response range than that observed with antibody-binding (Fab) fragment. Then, using a portable FPIA instrument, a VHH-based immunoassay of human IgG in a human serum certified reference material was demonstrated.

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A camelid VHH-based fluorescence polarization immunoassay for the detection of tetrabromobisphenol A in water

Abstract: Tetrabromobisphenol A (TBBPA) is a widely used flame retardant and is classified as an endocrine disruptor.

Cited by 7 publications

References 38 publications

BPA Endocrine Disruptor Detection at the Cutting Edge: FPIA and ELISA Immunoassays

BPA Endocrine Disruptor Detection at the Cutting Edge: FPIA and ELISA Immunoassays

Dual-Wavelength Fluorescence Polarization Immunoassay for Simultaneous Detection of Sulfonamides and Antibacterial Synergists in Milk

Noncompetitive Fluorescence Polarization Immunoassay for Protein Determination

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