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Strong and oriented conjugation of nanobodies onto magnetosomes for the development of a rapid immunomagnetic assay for the environmental detection of tetrabromobisphenol-A

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Abstract

Variable domain of heavy chain antibody (nanobody, Nb) derived from camelids is an efficient reagent in monitoring environmental contaminants. Oriented conjugates of Nbs and bacterial magnetic particles (BMPs) provide new tools for the high-throughput immunoassay techniques. An anti-tetrabromobisphenol-A (TBBPA) Nb genetically integrated with an extra cysteine residue at the C terminus was immobilized onto BMPs enclosed within the protein membrane, using a heterobifunctional reagent N-succinimidyl-3-(2-pyridyldithiol) propionate, to form a solid BMP-Nb complex. A rapid and sensitive enzyme-linked immunosorbent assay (ELISA) based on the combination of BMP-Nb and T5-horseradish peroxidase was developed for the analysis of TBBPA, with a total assay time of 30 min and a half-maximum signal inhibition concentration (IC50) of 1.04 ng/mL in PBS (pH 10, 10% methanol and 0.137 moL/L NaCl). This assay can even be performed in 100% methanol, with an IC50 value of 44.3 ng/mL. This assay showed quantitative recoveries of TBBPA from spiked canal water (114–124%) and sediment (109–113%) samples at 1.0–10 ng/mL (or ng/g (dw)). TBBPA residues determined by this assay in real canal water samples were below the limit of detection (LOD) and in real sediments were between <LOD and 23.4 ng/g (dw). The BMP-Nb-based ELISA shows promising application in environmental monitoring.

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Acknowledgements

This work was supported in part by the Project of the National Natural Science Foundation of China (21577170), Key Project of Inter-Governmental International Scientific and Technological Innovation Cooperation (2016YFE0108900), China and the National Institute of Environmental Health Sciences Superfund Research Program, P42ES04699, USA.

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Authors and Affiliations

  1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Resources and Environmental Sciences, China Agricultural University, No. 2 Yuan Ming Yuan West Road, Haidian District, Beijing, 100193, China

    Jinxin He, Kai Wang, Ji Li & Ting Xu

  2. Department of Microbiology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China

    Jiesheng Tian & Junjie Xu

  3. Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA, 95616, USA

    Shirley J. Gee & Bruce D. Hammock

  4. Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, 1955 East-West Road, Honolulu, HI, 96822, USA

    Qing X. Li

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  1. Jinxin He
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Correspondence to Ting Xu.

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He, J., Tian, J., Xu, J. et al. Strong and oriented conjugation of nanobodies onto magnetosomes for the development of a rapid immunomagnetic assay for the environmental detection of tetrabromobisphenol-A. Anal Bioanal Chem 410, 6633–6642 (2018). https://doi.org/10.1007/s00216-018-1270-9

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  • DOI: https://doi.org/10.1007/s00216-018-1270-9

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