Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs, Wuhan, China; Laboratory of Risk Assessment for Oilseeds Products, Ministry of Agriculture and Rural Affairs, Wuhan, China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan, China; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture and Rural Affairs, Wuhan, China; Laboratoire BAE-LBBM USR 3579, Université De Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex 66860, France.
Laboratoire BAE-LBBM USR 3579, Université De Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex 66860, France.
Food Chem. 2022 Jul 30;383:132598. doi: 10.1016/j.foodchem.2022.132598. Epub 2022 Mar 1.
This study aimed to devise a nontoxic electrochemical immunosensor to quantitatively determine aflatoxin M by chronoamperometry with novel anti-idiotypic nanobody-functionalized screen-printed carbon electrodes (SPCEs). Anti-idiotype nanobodies (AIdnb) were developed to replace the high toxic chemically synthesized antigen. AIdnb was immobilized on the surface of SPCE via covalent coupling as capture reagent. The functionalized SPCEs were followed by characterization using electrochemical impedance spectroscopy, fourier-transform infrared spectroscopy, transmission electron microscopy mapping, and atomic force microscopy. After optimizing experimental parameters, the assembled immunosensor exhibited a good linearity range of 0.25-5.0 ng/mL, with the limit of detection of 0.09 ng/mL. The immunosensor showed a satisfactory selectivity to AFM, without interference from analogs, including zearalenone, ochratoxin, and fumonisin B. For practical application, the developed immunosensor was validated using real spiked samples with the recovery range 82.0%-108.0% and relative standard deviation (RSD) 10.1%-13.0%, indicating that it could be used in milk samples.
本研究旨在通过计时安培法设计一种非毒性电化学免疫传感器，通过新型抗独特型纳米抗体功能化的丝网印刷碳电极（SPCE）定量测定黄曲霉毒素 M。开发了抗独特型纳米抗体（AIdnb）以替代高毒性的化学合成抗原。AIdnb 通过共价偶联固定在 SPCE 表面作为捕获试剂。通过电化学阻抗谱、傅里叶变换红外光谱、透射电子显微镜图谱和原子力显微镜对功能化的 SPCE 进行了表征。在优化实验参数后，组装的免疫传感器表现出 0.25-5.0 ng/mL 的良好线性范围，检测限为 0.09 ng/mL。该免疫传感器对 AFM 表现出令人满意的选择性，没有类似物（包括玉米赤霉烯酮、赭曲霉毒素和伏马菌素 B）的干扰。为了实际应用，使用实际添加的样品对开发的免疫传感器进行了验证，回收率范围为 82.0%-108.0%，相对标准偏差（RSD）为 10.1%-13.0%，表明它可用于牛奶样品。
Zotero 插件