数字农科院2.0

An Enhanced Ratiometric Fluorescence Immunosensor for Rapid Detection of AFB1 Based on the Dual-Enzyme Synergistic Co-Catalytic Effect of Natural Enzymes and Nanozymes

文献类型： 外文期刊

作者： Jing, Yinnan;Wang, Jiamin;Liu, Xuan;Li, Shurong;Yang, Qinkai;Wang, Mengnan;Fan, Xueyi;Ge, Guangsheng;Zheng, Kai;Wang, Jianlong;Tang, Xiaoqian;Zhang, Qi;Wang, Yanru

作者机构：

关键词： aflatoxin;anti-idiotypicnanobody;enzymesynergy;dual enzyme activity;ratiometric fluorescentsignal

期刊名称： JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

ISSN： 0021-8561

年卷期： 2025 年

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： Aflatoxin B1 (AFB1) is considered a critical analyte in food safety testing due to its widespread contamination and high toxicity. In this study, a ratiometric fluorescent immunosensor with synergistic catalysis by natural enzymes and nanozymes (N-N-RF-immunosensor) was constructed for the rapid detection of AFB1. Copper oxide and zeolitic imidazolate framework composite nanomaterials (CuO/ZIF-8 NCM) with peroxidase and oxidase activities were prepared. A ratiometric fluorescence signal was generated through the synergistic cocatalytic effect of the CuO/ZIF-8 NCM and alkaline phosphatase, in response to the competition interaction between the anti-idiotypic nanobody-alkaline phosphatase fusion protein (Nb2-5-AP) and AFB1. The assay could detect aflatoxins within 140 min, exhibiting a half maximal inhibitory concentration (IC50) of 0.032 ng/mL, a limit of detection (LOD) of 2.4 pg/mL, and a linear range of 0.0024-1.043 ng/mL. Compared with the traditional enzyme-linked immunosorbent assay (ELISA), the sensitivity was enhanced 25-fold, while the detection parameter was shortened by 1/3. Furthermore, the immunosensor was successfully applied in the actual samples, with recoveries ranging from 81 to 127%. The N-N-RF-immunosensor shows great potential for rapid and sensitive detection of aflatoxins.

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