Target-induced catalytic hairpin assembly formation of functional Y-junction DNA structures for label-free and sensitive electrochemical detection of human serum proteins
Target-induced catalytic hairpin assembly formation of functional Y-junction DNA structures for label-free and sensitive electrochemical detection of human serum proteins
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靶标诱导催化发夹组装形成功能性 Y 接头 DNA 结构，用于人血清蛋白的无标记且灵敏的电化学检测
DOI：
10.1016/j.snb.2016.12.131
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发表时间：
2017-06
期刊：
影响因子：
--
通讯作者：
Yuan Ruo
中科院分区：
文献类型：
--
作者：
Jiang Bingying;Li Fangzhen;Yan Cuiyun;Xie Jiaqing;Xiang Yun;Yuan Ruo
构建简单、高灵敏度的适体传感器将极大地促进蛋白质的监测。在这项工作中，我们建立了一种简便的、无标记的蛋白质诱导的催化发夹组装信号放大方法，用于基于适体的灵敏的人血清凝血酶的电化学检测。靶凝血酶与固定在传感器电极上的含适配子的发夹探针结合，并触发另外两个发夹的催化组装，形成许多G-四链Y-连接DNA结构。这些G-四链体结构进一步与氯化高铁血红素结合形成G-四链体/氯化高铁血红素复合体，然后进行电化学测量，并产生大量放大的电流输出，用于无标记和灵敏的凝血酶检测。所研制的适配传感器对凝血酶的检测动态范围为0.01-1.0nM，检测下限为6.0nM。该传感器还可以将目标凝血酶与其他非目标蛋白区分开来，并显示出检测人血清中凝血酶的良好结果。随着凝血酶适配子传感器的成功构建，我们的信号放大方法可以通过选择合适的适配子/配体对来检测其他靶分子。
The construction of simple and highly sensitive aptasensors will significantly advance the monitoring of proteins. In this work, we have developed a convenient and label-free protein-induced catalytic hairpin assembly signal amplification approach for aptamer-based sensitive electrochemical detection of thrombin in human serums. The target thrombin binds the aptamer-containing hairpin probes immobilized on the sensor electrodes and triggers catalytic assembly of other two hairpins to form many G-quadruplex Y-junction DNA structuresin situ. These G-quadruplex structures further associate with hemin to form G-quadruplex/hemin complexes, which are then subject to electrochemical measurement and generate substantially amplified current output for label-free and sensitive detection of thrombin. The developed aptasensor shows a dynamic range of 0.01–1.0 nM and a detection limit of 6.0 p.m for thrombin detection. The sensor can also discriminate the target thrombin against other non-target proteins and exhibits promising results for detecting thrombin in human serums. With the successful construction of the thrombin aptasensor, our signal amplified method can be potentially applied to detect other target molecules by choosing appropriate aptamer/ligand pairs.
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