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An ultrasensitive, homogeneous fluorescence quenching immunoassay integrating separation and detection of aflatoxin M1 based on magnetic graphene composites

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Abstract

A homogeneous fluorescence quenching immunoassay is described for simultaneous separation and detection of aflatoxin M1 (AFM1) in milk. The novel assay relies on monoclonal antibody (mAb) functionalized Fe3O4 decorated reduced-graphene oxide (rGO-Fe3O4-mAb) as both capture probe and energy acceptor, combined with tetramethylrhodamine cadaverine-labeled aflatoxin B1 (AFB1-TRCA) as the energy donor. In the assay, AFB1-TRCA binds to rGO-Fe3O4-mAb in the absence of AFM1, quenching the fluorescence of TRCA by resonance energy transfer. Significantly, the immunoassay integrates sample preparation and detection into a single step, by using magnetic graphene composites to avoid washing and centrifugation steps, and the assay can be completed within 10 min. Under optimized conditions, the visual and quantitative detection limits of the assay for AFM1 were 50 and 3.8 ng L−1, respectively, which were significantly lower than those obtained by fluorescence polarization immunoassay using the same immunoreagents. Owing to its operation and highly sensitivity, the proposed assay provides a powerful tool for the detection of AFM1.

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Funding

This study was supported by Natural Science Foundation of China (No. 31802249), Science and Technology Key Research Project of Henan Provincial Education Department of China (No. 20A550009) and Key Scientific and Technological Project of Henan Province Department of China (202102310303 and 202102110103).

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Authors and Affiliations

  1. Henan Province Engineering Research Center for Food Safety Control of Processing and Circulation, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou, 450002, Henan, China

    Xiya Zhang, Lianjun Song, Xianqing Huang, Yu Li, Mingwu Qiao, Wanjing Liu, Tongtong Zhang, Yuchen Qi, Wuzhou Wang & Yexuan Mao

  2. The Third Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, China

    Xijie Zhang

  3. Beijing Key Laboratory of Detection Technology for Animal–Derived Food Safety, Beijing Laboratory of Food Quality and Safety, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China

    Xuezhi Yu, Leina Dou, Huijuan Yang & Zhanhui Wang

  4. College of Food and Drug, Luoyang Normal University, Luoyang, 471934, Henan, China

    Liye Wang

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  1. Xiya Zhang
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Correspondence to Yexuan Mao.

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Supplementary information

It is mainly contained preparation of scFv-1D3, homology modeling of scFv-1D3, molecular recognition of scFv-1D3 and development of the FPIA for AFM1 in milk samples. The details are available free of charge via the Internet at http://...

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Zhang, X., Zhang, X., Song, L. et al. An ultrasensitive, homogeneous fluorescence quenching immunoassay integrating separation and detection of aflatoxin M1 based on magnetic graphene composites. Microchim Acta 188, 59 (2021). https://doi.org/10.1007/s00604-021-04715-2

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