Dang Jie, Guo Zhihui, Zheng Xingwang
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University , Xi'an 710062, P.R. China.
Anal Chem. 2014 Sep 16;86(18):8943-50. doi: 10.1021/ac5017584. Epub 2014 Aug 29.
In this work, a label-free and sensitive electrogenerated chemiluminescence (ECL) aptasensing scheme for K(+) was developed based on G-rich DNA aptamer and chitosan/Ru(bpy)3(2+)/silica (CRuS) nanoparticles (NPs)-modified glass carbon electrode. This ECL aptasensing approach has benefited from the observation that the G-rich DNA aptamer at the unfolded state showed more ECL enhancing signal at CRuS NPs-modified electrode than the binding state with K(+), which folds into G-quadruplex structure. As such, the decreasing ECL signals could be used to detect K(+). Compared to other aptasensing K(+) approaches previously reported, the proposed ECL sensing scheme is a label-free aptasensing strategy, which eliminates the labeling, separation, and immobilization steps, and behaves in a simple, low-cost way. More importantly, because the proposed ECL sensing mechanism utilizes the nanosized ECL active CRuS NPs to sense the nanoscale conformation change from the aptamer binding to target, it is specific. In addition, due to the great conformation changes of the aptamer's G-bases on CRuS NPs and the excellent ECL enhancing effect of guanine bases to the Ru(bpy)3(2+) ECL reaction, a 0.3 nM detection limit for K(+) was achieved with the proposed ECL method. On the basis of these advantages, the proposed ECL aptasensing method was also successfully used to detect K(+) in colorectal cancer cells.
在本研究中,基于富含鸟嘌呤的DNA适配体以及壳聚糖/三联吡啶钌(Ⅱ)/二氧化硅(CRuS)纳米颗粒修饰的玻碳电极,开发了一种用于检测钾离子的无标记且灵敏的电化学发光(ECL)适配体传感方案。这种ECL适配体传感方法得益于以下发现:处于未折叠状态的富含鸟嘌呤的DNA适配体在CRuS纳米颗粒修饰电极上比与钾离子结合形成G-四链体结构时显示出更强的ECL增强信号。因此,ECL信号的降低可用于检测钾离子。与先前报道的其他检测钾离子的适配体传感方法相比,所提出的ECL传感方案是一种无标记的适配体传感策略,它省去了标记、分离和固定步骤,操作简单且成本低廉。更重要的是,由于所提出的ECL传感机制利用纳米尺寸的ECL活性CRuS纳米颗粒来感知适配体与靶标结合引起的纳米级构象变化,所以具有特异性。此外,由于适配体的鸟嘌呤碱基在CRuS纳米颗粒上发生了巨大的构象变化,且鸟嘌呤碱基对三联吡啶钌(Ⅱ)的ECL反应具有优异的增强效果,所提出的ECL方法对钾离子的检测限达到了0.3 nM。基于这些优点,所提出的ECL适配体传感方法还成功用于检测结肠癌细胞中的钾离子。