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Flow cytometric immunoassay for aflatoxin B1 using magnetic microspheres encoded with upconverting fluorescent nanocrystals

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Abstract

The authors describe a flow cytometric immunoassay for aflatoxin B1 (AFB1). It has three distinct features: (a) Magnetic microspheres encoded with upconverting nanocrystals (UCNMMs) are used as fluorescent labels. These have the advantage of non-overlapping spectra and lacking crosstalk between the encoding signal and reporter signal via the low-energy near-infrared (NIR) light excitation; (b) phycoerythrin-labeled secondary antibodies are used to amplify the reporter signal; (c) The use of magnetic nanoparticles facilitates the rapid separation and specific purification of the analyte (AFB1). This assay has a detection limit of 9 pg·mL−1 and a broad working range for AFB1, requires a 50 μL sample only, and can be completed within 2 h with good accuracy and high reproducibility. It is perceived that such multifluorescent UCNMMs, whose color depends on the kind of dopants (Yb, Er, Tm, Mn) in the NaYF4 host lattice, represent a promising tool for the analysis of mycotoxins and other analytes.

Schematic of the UCNMM-based indirective competitive immunoassay for AFB1 using the flow cytometric analysis (FCA) technology. The UCNMMs are prepared by doping the upconversion nanocrystals and magnetic nanoparticles inside the mesoporous polystyrene microspheres as the self-healing encapsulation strategy.

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Acknowledgements

The authors gratefully acknowledge National Natural Science Foundation of China (31401578, 51373117, 51303126, 51573128 and 81402575), Tianjin Natural Science Foundation (13JCZDJC33200 and 15JCQNJC03100).

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Authors and Affiliations

  1. College of Life Sciences, Tianjin University, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Tianjin, 300072, People’s Republic of China

    Ying Zhang, Yajuan Liu, Jin Chang & Hanjie Wang

  2. The National Center of Supervision and Inspection for Quality of Food, Tianjin Product Quality Inspection Technology Research Institute, Tianjin, 300384, People’s Republic of China

    Zhenyu Liao

  3. College of Life Sciences, Nankai University, Tianjin, 300071, People’s Republic of China

    Yajuan Wan

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  1. Ying Zhang
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  2. Zhenyu Liao
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  3. Yajuan Liu
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  5. Jin Chang
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Correspondence to Zhenyu Liao or Hanjie Wang.

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Zhang, Y., Liao, Z., Liu, Y. et al. Flow cytometric immunoassay for aflatoxin B1 using magnetic microspheres encoded with upconverting fluorescent nanocrystals. Microchim Acta 184, 1471–1479 (2017). https://doi.org/10.1007/s00604-017-2116-4

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  • DOI: https://doi.org/10.1007/s00604-017-2116-4

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