Peng Huaping, Huang Zhongnan, Wu Weihua, Liu Mingkai, Huang Kaiyuan, Yang Yu, Deng Haohua, Xia Xinghua, Chen Wei
Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, Department of Pharmaceutical Analysis , Fujian Medical University , Fuzhou 350004 , China.
School of Chemistry & Materials Science, Jiangsu Key Laboratory of Green Synthetic Chemistry for Functional Materials , Jiangsu Normal University , Xuzhou 221116 , China.
ACS Appl Mater Interfaces. 2019 Jul 10;11(27):24812-24819. doi: 10.1021/acsami.9b08819. Epub 2019 Jun 26.
This report outlines a versatile high-performance electrochemiluminescence (ECL) enzyme-linked immunosorbent assay (ELISA) platform, which combines the merits of high-quantum-yield Au nanocluster (AuNC) probe-based ECL technology, the efficient ECL-resonance energy-transfer (ECL-RET) strategy, and highly sensitive and specific ELISA technology. The ECL detection procedure was performed on a recyclable MnO/AuNC-modified glassy carbon electrode interface by taking advantage of the ECL-RET between the AuNC probe and MnO nanomaterials (NMs) to quench the ECL intensity. The etching of MnO NMs by the product of ALP-based ELISA recovers the ECL signal. Notably, the ELISA process and the ECL detection procedure in this system are independent. Thus, the ECL-ELISA system can effectively avoid the influence of complex biological samples, and the ECL efficiency of the AuNC probe can be used readily. As demonstrated on TNF-α, because of the abovementioned characteristics, the ECL-ELISA platform presented an extremely wide dynamic range, with a detection limit of 2 orders lower than ELISA. Moreover, the system was also applicable for ultrahigh sensitive detection of various disease-related proteins and able to detect trace biomarkers in real serum samples. Therefore, this multifunctional ECL assay platform is versatile, facile, ultrasensitive, recyclable, and sufficiently straightforward for trace biomarker detection in complex biological samples. This approach not only enriches the foundational study of ECL devices but also greatly expands the potential application of ECL sensors in biological testing and clinical high-throughput diagnosis.
本报告概述了一种多功能高性能电化学发光（ECL）酶联免疫吸附测定（ELISA）平台，该平台结合了基于高量子产率金纳米簇（AuNC）探针的ECL技术、高效的ECL共振能量转移（ECL-RET）策略以及高灵敏度和特异性的ELISA技术的优点。利用AuNC探针与MnO纳米材料（NMs）之间的ECL-RET淬灭ECL强度，在可回收的MnO/AuNC修饰玻碳电极界面上进行ECL检测过程。基于碱性磷酸酶（ALP）的ELISA产物对MnO NMs的蚀刻恢复了ECL信号。值得注意的是，该系统中的ELISA过程和ECL检测过程是独立的。因此，ECL-ELISA系统可以有效避免复杂生物样品的影响，并且可以方便地使用AuNC探针的ECL效率。如在肿瘤坏死因子-α（TNF-α）上所证明的，由于上述特性，ECL-ELISA平台呈现出极宽的动态范围，检测限比ELISA低2个数量级。此外，该系统还适用于各种疾病相关蛋白的超高灵敏度检测，并能够检测真实血清样品中的痕量生物标志物。因此，这种多功能ECL测定平台通用、简便、超灵敏、可回收，并且对于复杂生物样品中的痕量生物标志物检测足够直接。这种方法不仅丰富了ECL器件的基础研究，而且极大地扩展了ECL传感器在生物检测和临床高通量诊断中的潜在应用。
