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 1 ; Wang, Jiamin 1 ; Liu, Xuan 1 ; Li, Shurong 1 ; Yang, Qinkai 1 ; Wang, Mengnan 1 ; Fan, Xueyi 1 ; Ge, Guangsheng 1 ; Zheng, Kai 1 ; Wang, Jianlong 1 ; Tang, Xiaoqian 2 ; Zhang, Qi 2 ; Wang, Yanru 1 ;
作者机构： 1.Northwest A&F Univ, Coll Food Sci & Engn, Yangling 712100, Shaanxi, Peoples R China
2.Chinese Acad Agr Sci, Oil Crops Res Inst, Wuhan 430062, Peoples R China
关键词： aflatoxin; anti-idiotypicnanobody; enzymesynergy; dual enzyme activity; ratiometric fluorescentsignal
期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：6.2； 五年影响因子：6.4 ）
ISSN： 0021-8561
年卷期： 2025 年 73 卷 14 期
页码：
收录情况： SCI
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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