Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China; Department of Orthopaedic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Biosens Bioelectron. 2019 Apr 1;130:88-94. doi: 10.1016/j.bios.2019.01.023. Epub 2019 Jan 18.
Herein, tyramide signal amplification (TSA)-based electrochemical immunosensor was exploited for highly sensitive detection of CTCs. In this immunosensor, the nucleolin-targeting aptamer AS1411 (CP) was used to specifically capture tumor cells, and a TSA-based signal enhancement system consisting of Pt NPs@HRP@CP composite as catalytic probe and tyramine functionalized infinite coordination polymer (ICPs@Tyr) as electroactive signal tag was applied to improve the detection sensitivity. Using HeLa cell as the model CTCs, after a sandwich reaction, CP-HeLa-Pt NPs@HRP@CP bioconjugates were formed on the electrode. Millions of ICPs@Tyr could be layer-by-layer deposited onto the target cell membrane by the catalysis of Pt NPs@HRP@CP. The developed immunorsensor could detect HeLa cell with a wide dynamic range from 2 to 2 × 10 cells/mL and a detection limit of 2 cells/mL. Most importantly, the amperometric immunosensor was successfully applied to discriminate tumor cells from white blood cells, exhibiting high specificity and sensitivity. In conclusion, this work demonstrates that the TSA-based signal enhancement system might be a potential alternative tool for the electrochemical measurement of trace amounts of CTCs in clinical diagnosis.
在此,我们利用酪胺信号放大(TSA)电化学免疫传感器来高灵敏度地检测循环肿瘤细胞(CTC)。在该免疫传感器中,使用核仁素靶向适体 AS1411(CP)特异性捕获肿瘤细胞,并且应用基于 TSA 的信号增强系统,该系统由 Pt NPs@HRP@CP 复合作为催化探针和酪胺功能化的无限配位聚合物(ICPs@Tyr)作为电活性信号标记物,以提高检测灵敏度。使用 HeLa 细胞作为模型 CTC,在三明治反应后,CP-HeLa-Pt NPs@HRP@CP 生物缀合物形成在电极上。Pt NPs@HRP@CP 的催化作用可将数百万个 ICPs@Tyr 逐层沉积到靶细胞膜上。所开发的免疫传感器能够检测到动态范围为 2 至 2×10 个/mL 的 HeLa 细胞,检测限为 2 个/mL。最重要的是,该电化学免疫传感器成功地用于区分肿瘤细胞和白细胞,表现出高特异性和灵敏度。总之,这项工作表明,基于 TSA 的信号增强系统可能是临床诊断中痕量 CTC 电化学测量的潜在替代工具。