MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China.
Anal Chem. 2023 Feb 14;95(6):3434-3441. doi: 10.1021/acs.analchem.2c05164. Epub 2023 Jan 31.
In this study, we developed, for the first time, a novel dry chemistry-based bipolar electrochemiluminescence (ECL) immunoassay device for point-of-care testing (POCT) of Alzheimer-associated neuronal thread protein (AD7c-NTP), where the ECL signals were automatically collected and analyzed after the sample and buffer solutions were manually added onto the immunosensor. The proposed immunoassay device contains an automatic ECL analyzer and a dry chemistry-based ECL immunosensor fabricated with a screen-printed fiber material-based chip and a three-dimensional (3D)-printed shell. Each pad of the fiber material-based chip was premodified with certain reagents for immunoreaction and then assembled to form the ECL immunosensor. The self-enhanced ECL of the Ru(II)-poly-l-lysine complex and the lateral flow fiber material-based chip make the addition of coreactants and repeated flushing unnecessary. Only the sample and buffer solutions are added to the ECL immunosensor, and the process of ECL detection can be completed in about 6 min using the proposed automatic ECL analyzer. Under optimized conditions, the linear detection range for AD7c-NTP was 1 to 10 pg/mL, and the detection limit was 0.15 pg/mL. The proposed ECL immunoassay device had acceptable selectivity, stability, and reproducibility and had been successfully applied to detect AD7c-NTP levels in human urine. In addition, the accurate detection of AD7c-NTP and duplex detection of AD7c-NTP and apolipoprotein E ε4 gene were also validated. It is believed that the proposed ECL immunoassay device may be a candidate for future POCT applications.
在本研究中，我们首次开发了一种基于干化学的新型双极电化学发光（ECL）免疫分析装置，用于即时检测（POCT）阿尔茨海默病相关神经元丝蛋白（AD7c-NTP），其中在手动将样品和缓冲溶液添加到免疫传感器上后，自动收集和分析 ECL 信号。所提出的免疫分析装置包含自动 ECL 分析仪和基于干化学的 ECL 免疫传感器，该免疫传感器由基于丝网印刷纤维材料的芯片和三维（3D）打印外壳制成。纤维材料基芯片的每个垫都经过特定的免疫反应试剂预修饰，然后组装形成 ECL 免疫传感器。Ru（II）-聚-L-赖氨酸配合物的自增强 ECL 和横向流动纤维材料基芯片使得无需添加共反应物和重复冲洗。只需将样品和缓冲溶液添加到 ECL 免疫传感器中，使用所提出的自动 ECL 分析仪，大约 6 分钟即可完成 ECL 检测过程。在优化条件下，AD7c-NTP 的线性检测范围为 1 至 10 pg/mL，检测限为 0.15 pg/mL。所提出的 ECL 免疫分析装置具有可接受的选择性、稳定性和重现性，并已成功应用于检测人尿液中的 AD7c-NTP 水平。此外，还验证了 AD7c-NTP 的准确检测和 AD7c-NTP 与载脂蛋白 E ε4 基因的双重检测。相信所提出的 ECL 免疫分析装置可能是未来 POCT 应用的候选者。
