Sensitivity enhancement of paper-based sandwich immunosensor via nanobody immobilization instead of IgG antibody, taking aflatoxingenetic fungi as an analyte example

文献类型： 外文期刊

第一作者： Yan, Honglin

作者： Yan, Honglin;Fu, Jiayun;Tang, Xiaoqian;Wang, Du;Zhang, Qi;Li, Peiwu;Zhang, Qi;Li, Peiwu;Zhang, Qi

作者机构：

关键词： Nanobody; Time-resolved fluorescence immunoassay (TRFIA); Immunosensor; Aflatoxingenetic fungi; Macromolecular hazard materials

期刊名称：SENSORS AND ACTUATORS B-CHEMICAL （ 影响因子：9.221； 五年影响因子：7.676 ）

ISSN：

年卷期： 2022 年 373 卷

页码：

收录情况： SCI

摘要： Aflatoxingenetic fungi producing highly toxic and carcinogenic mycotoxins and threatening human health widely exist in our living environment and food. Taking aflatoxingenetic fungi as an analyte example, here we report a way to improve the sensitivity of paper-based sandwich immunosensor for macromolecule. With a biomarker-specific nanobody and polyclonal antibody (IgG), two modes of paper-based immunosensors were constructed, immobilization nanobody mode and immobilization IgG mode. It was found that the sensitivity of the immobilization nanobody mode was at least 500 times higher than that of the immobilization IgG mode. The reasons for the large difference in sensitivity were also theoretically. Under the optimized conditions, the immobilization nanobody mode sensitivity reached 0.035 mu g/mL aflatoxingenetic mycelia, each assay was finished within 15 min, and the spiked recoveries of aflatoxingenetic mycelium reference in peanuts were 77.0 similar to 91.6%. And then, two important application cases were successfully demonstrated to identify the abundance of aflatoxingenetic fungi in market peanuts and peanut rhizosphere soil for environmental safety evaluation. This is the first report of time-resolved paper-based sandwich immunosensor for aflatoxingenetic fungi with nanobody immobilization, and the first sensitivity comparison between nanobody immobilization and IgG immobilization, which provides a new approach to develop highly sensitive immunosensor for macromolecular hazard materials.

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