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A sensitive electrochemical immunosensing interface for label-free detection of aflatoxin B1 by attachment of nanobody to MWCNTs-COOH@black phosphorene

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Abstract

A label-free electrochemical immunosensor has advantages of real-time and rapid detection, but it is weak in detection of small molecular toxins such as aflatoxin B1 (AFB1). The greatest obstacle to achieving this is that small molecules bound to a common immunosensing interface cannot interfere with electron transfer effectively and the detection signal is so weak. Therefore, a sensitive electrochemical immunosensing interface for small molecules is urgently needed. Here, we employed functionalized black phosphorene (BP) as electrode modification materials and anti-AFB1 nanobody (Nb) as a biorecognition element to construct a very sensitive immunosensing interface towards small molecular AFB1. The BP functionalized by carboxylic multi-walled carbon nanotubes (MWCNTs-COOH) via P–C bonding behaved with a satisfactory stability and good catalytic performance for the ferricyanide/ferrocyanide probe, while the small-sized Nb showed good compatibility with the functionalized BP and also had less influence on electron transfer than monoclonal antibody (mAb). Expectedly, the as-prepared immunosensing interface was very sensitive to AFB1 detection by differential pulse voltammetry (DPV) in a redox probe system. Under optimized conditions, a linear range from 1.0 pM to 5.0 nM and an ultralow detection limit of 0.27 pM were obtained. Additionally, the fabricated immunosensor exhibited satisfactory stability, specificity, and reproducibility. The strategy proposed here provides a more reliable reference for label-free sensing of small molecules in food samples.

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Funding

This work was supported by the National Natural Science Foundation of China (31660492, 32060575, 32160602, 31860260) and the Natural Science Foundation of Jiangxi Province (20192ACBL20019, 20181BAB204017, 20171BAB214038).

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Authors and Affiliations

  1. Research Center of Mycotoxin, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045, People’s Republic of China

    Xue Zhang, Xiaoning Liao, Yongfa Wu & Dan Wang

  2. Jiangxi Key Laboratory for Postharvest Technology and Nondestructive Testing of Fruits & Vegetables/Collaborative Innovation Center of Postharvest Key Technology and Quality Safety of Fruits &Vegetables in Jiangxi Province, Jiangxi Agricultural University, Nanchang, 330045, Jiangxi, China

    Xue Zhang, Xiaoning Liao, Yongfa Wu & Wanming Xiong

  3. Key Lab for Agro-Product Processing and Quality Control of Nanchang City, College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang, 330045, Jiangxi, China

    Juan Du, Wuying Yang & Dan Wang

  4. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, Jiangxi, China

    Zhui Tu

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  1. Xue Zhang
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Xue Zhang and Xiaoning Liao contributed equally.

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Zhang, X., Liao, X., Wu, Y. et al. A sensitive electrochemical immunosensing interface for label-free detection of aflatoxin B1 by attachment of nanobody to MWCNTs-COOH@black phosphorene. Anal Bioanal Chem 414, 1129–1139 (2022). https://doi.org/10.1007/s00216-021-03738-1

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