State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.
College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001, China.
Anal Chem. 2021 Jan 19;93(2):1200-1208. doi: 10.1021/acs.analchem.0c04628. Epub 2020 Dec 10.
Engineered nanopipette tools have recently emerged as a powerful approach for electrochemical nanosensing, which has major implications in both fundamental biological research and biomedical applications. Herein, we describe a generic method of target-triggered assembly of aptamers in a nanopipette for nanosensing, which is exemplified by sensitive and rapid electrochemical single-cell analysis of adenosine triphosphate (ATP), a ubiquitous energy source in life and important signaling molecules in many physiological processes. Specifically, a layer of thiolated aptamers is immobilized onto a Au-coated interior wall of a nanopipette tip. With backfilled pairing aptamers, the engineered nanopipette is then used for probing intracellular ATP via the ATP-dependent linkage of the split aptamers. Due to the higher surface charge density from the aptamer assembly, the nanosensor would exhibit an enhanced rectification signal. Besides, this ATP-responsive nanopipette tool possesses excellent selectivity and stability as well as high recyclability. This work provides a practical single-cell nanosensor capable of intracellular ATP analysis. More generally, integrated with other split recognition elements, the proposed mechanism could serve as a viable basis for addressing many other important biological species.
近年来，工程纳米管工具作为电化学纳米传感的一种强大方法已经出现，这对基础生物学研究和生物医学应用都具有重大意义。在此，我们描述了一种在纳米管中靶触发组装适体的通用方法，该方法通过敏感和快速的电化学单细胞分析三磷酸腺苷（ATP）为例进行了说明，ATP 是生命中普遍存在的能量源，也是许多生理过程中的重要信号分子。具体来说，一层巯基化适体被固定在纳米管尖端的 Au 涂层内壁上。用回充配对适体，然后通过分裂适体的 ATP 依赖性连接，使用工程纳米管来探测细胞内的 ATP。由于组装适体的表面电荷密度更高，纳米传感器将表现出增强的整流信号。此外，这种对 ATP 有响应的纳米管工具具有出色的选择性、稳定性和高可回收性。这项工作提供了一种实用的单细胞纳米传感器，能够进行细胞内 ATP 分析。更一般地说，与其他分裂识别元件结合，所提出的机制可以作为解决许多其他重要生物物质的可行基础。
