National and Local United Engineering Lab of Druggability and New Drugs Evaluation, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou, 510006, Guangdong, China; Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou, 510006, Guangdong, China.
Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou, 510006, Guangdong, China.
Biosens Bioelectron. 2024 Dec 15;266:116717. doi: 10.1016/j.bios.2024.116717. Epub 2024 Aug 28.
Epithelial cell adhesion molecule (EpCAM), which is overexpressed in breast cancer cells and participates in cell signaling, migration, proliferation, and differentiation, has been utilized as a biomarker for cancer diagnosis and therapeutic prognosis. Here, a dual-signal readout nonenzymatic aptasensor is fabricated for the evaluation of EpCAM at the level of three breast cancer cell lines. The central principle of this enzyme-free aptasensor is the use of double hook-type aptamers (SYL3C and SJ3C2)-functionalized magnetic iron oxide (FeO) as capture probes and quasi-CoFe prussian blue analogs (QCoFe PBAs) as nonenzymatic signal probes for colorimetric and electrochemical analysis. Following ligand detachment, the CoFe PBA was transformed to QCoFe PBA (calcined at 350 °C for 1 h), with its metal active sites exposed by controllable pyrolysis. We found that the enhanced sensitivity was attributed to the resonance effect of QCoFe PBA with the remarkable enzymatic properties. The dual-signal readout nonenzymatic aptasensor exhibited limits of detection for EpCAM as low as 0.89 pg mL and 0.24 pg mL, within a wide linear range from 0.001 to 100 ng mL, respectively. We successfully employed this nonenzymatic aptasensor for monitoring EpCAM expression in three breast cancer cell lines, which provides an economical and robust alternative to costly and empirical flow cytometry. The dual-signal readout nonenzymatic aptasensor provides rapid, robust, and promising technological support for the accurate management of tumors.
上皮细胞黏附分子（EpCAM）在乳腺癌细胞中过度表达，并参与细胞信号转导、迁移、增殖和分化，已被用作癌症诊断和治疗预后的生物标志物。在这里，构建了一种双信号读出无酶适体传感器，用于评估三种乳腺癌细胞系中 EpCAM 的水平。该无酶适体传感器的核心原理是使用双钩型适体（SYL3C 和 SJ3C2）功能化的磁性氧化铁（FeO）作为捕获探针和类普鲁士蓝纳米酶（QCoFe PBAs）作为比色和电化学分析的非酶信号探针。配体脱离后，CoFe PBA 被转化为 QCoFe PBA（在 350°C 下煅烧 1 小时），其金属活性位通过可控热解暴露。我们发现，增强的灵敏度归因于 QCoFe PBA 的共振效应及其显著的酶学性质。双信号读出无酶适体传感器对 EpCAM 的检测限低至 0.89pg/mL 和 0.24pg/mL，在 0.001 至 100ng/mL 的宽线性范围内分别具有较高的灵敏度。我们成功地将这种无酶适体传感器用于监测三种乳腺癌细胞系中的 EpCAM 表达，为昂贵且经验性的流式细胞术提供了一种经济且强大的替代方案。双信号读出无酶适体传感器为肿瘤的精确管理提供了快速、稳健、有前景的技术支持。
