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用于灵敏电化学生物传感的简单可调节 DNA 二聚体纳米器件来排列级联酶。

Simple and Regulable DNA Dimer Nanodevice to Arrange Cascade Enzymes for Sensitive Electrochemical Biosensing.

机构信息

Key Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, P. R. China.

出版信息

Anal Chem. 2020 Oct 20;92(20):14197-14202. doi: 10.1021/acs.analchem.0c03396. Epub 2020 Sep 30.

DOI:10.1021/acs.analchem.0c03396
PMID:32938178
Abstract

Herein, a simple and regulable DNA dimer nanodevice was obtained by the assembly of two hairpin DNA monomers of H1 and H2 to control the distance between model enzymes horseradish peroxidase (HRP) and glucose oxidase (GOx) for sensitive electrochemical detection of microRNA. In detail, auto-terminated DNA polymerization reaction was designed on H1 and H2 monomers that decorated with HRP and GOx, respectively, to produce two half-released DNA monomers, which were spontaneously hybridized to each other, thereby obtaining a DNA dimer nanodevice with a rigid dsDNA linker between two DNA monomers. By varying the length of the dsDNA linker on the DNA dimer nanodevice, the distance between GOx and HRP had been regulated to the optimum and the most efficient enzyme cascade reaction was acquired for constructing a sensitive electrochemical microRNA-21 biosensor with a detection limit of 0.03 pM. In summary, the proposed DNA dimer nanodevice avoided the disadvantages of poor biocompatibility and controllability originated from traditional scleroid scaffolds and showed obvious advantages in terms of better assembly yield than previous complicated DNA scaffolds, which provided a novel strategy for developing a high-efficiency enzyme cascade catalytic system and showed great potential in other clinical diagnosis and bioanalysis application.

摘要

本文通过组装两个发夹 DNA 单体 H1 和 H2,获得了一种简单且可调的 DNA 二聚体纳米器件,以控制模型酶辣根过氧化物酶 (HRP) 和葡萄糖氧化酶 (GOx) 之间的距离,用于灵敏电化学检测 microRNA。具体来说,在分别修饰有 HRP 和 GOx 的 H1 和 H2 单体上设计了自动终止的 DNA 聚合反应,产生两个半释放的 DNA 单体,它们自发杂交,从而获得具有刚性 dsDNA 连接子的 DNA 二聚体纳米器件在 DNA 二聚体纳米器件上改变 dsDNA 连接子的长度,可以调节 GOx 和 HRP 之间的距离,达到最佳状态,并获得最有效的酶级联反应,从而构建具有检测限为 0.03 pM 的灵敏电化学 microRNA-21 生物传感器。总之,所提出的 DNA 二聚体纳米器件避免了传统刚性支架固有的生物相容性和可控性差的缺点,并且与以前复杂的 DNA 支架相比,显示出更好的组装产率方面的明显优势,为开发高效酶级联催化系统提供了一种新策略,并在其他临床诊断和生物分析应用中显示出巨大的潜力。

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