Low-Noise Solid-State Nanopore Enhancing Direct Label-Free Analysis for Small Dimensional Assemblies Induced by Specific Molecular Binding,ACS Applied Materials & Interfaces

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Low-Noise Solid-State Nanopore Enhancing Direct Label-Free Analysis for Small Dimensional Assemblies Induced by Specific Molecular Binding
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2021-01-21 , DOI: 10.1021/acsami.0c20359
Ruiping Wu _{1,

2} , Yesheng Wang _{1,

2} , Zhentong Zhu ₃ , Chunmiao Yu _{1,

2} , Huan Li _{1,

2} , Bingling Li _{1,

2} , Shaojun Dong _{1,

2}

Affiliation

Solid-state nanopores show special potential as a new single-molecular characterization for nucleic acid assemblies and molecular machines. However, direct recognition of small dimensional species is still quite difficult due the lower resolution compared with biological pores. We recently reported a very efficient noise-reduction and resolution-enhancement mechanism via introducing high-dielectric additives (e.g., formamide) into conical glass nanopore (CGN) test buffer. Based on this advance, here, for the first time, we apply a bare CGN to directly recognize small dimensional assemblies induced by small molecules. Cocaine and its split aptamer (Capt assembly) are chosen as the model set. By introducing 20% formamide into CGN test buffer, high cocaine-specific distinguishing of the 113 nt Capt assembly has been realized without any covalent label or additional signaling strategies. The signal-to-background discrimination is much enhanced compared with control characterizations such as gel electrophoresis and fluorescence resonance energy transfer (FRET). As a further innovation, we verify that low-noise CGN can also enhance the resolution of small conformational/size changes happening on the side chain of large dimensional substrates. Long duplex concatamers generated from the hybridization chain reaction (HCR) are selected as the model substrates. In the presence of cocaine, low-noise CGN has sensitively captured the current changes when the 26 nt aptamer segment is assembled on the side chain of HCR duplexes. This paper proves that the introduction of the low-noise mechanism has significantly improved the resolution of the solid-state nanopore at smaller and finer scales and thus may direct extensive and deeper research in the field of CGN-based analysis at both single-molecular and statistical levels, such as molecular recognition, assembly characterization, structure identification, information storage, and target index.

中文翻译：

低噪声固态纳米孔增强了由特定分子结合引起的小尺寸组装体的直接无标签分析

固态纳米孔作为核酸组装和分子机器的一种新的单分子表征具有特殊的潜力。但是，由于与生物孔相比分辨率较低，因此直接识别小尺寸物种仍然非常困难。我们最近报道了一种通过将高介电添加剂（例如甲酰胺）引入锥形玻璃纳米孔（CGN）测试缓冲液中的非常有效的降噪和分辨率增强机制。基于此进展，我们首次在此应用裸CGN直接识别由小分子诱导的小尺寸组装。选择可卡因及其拆分适体（Capt装配体）作为模型集。通过向CGN测试缓冲液中引入20％甲酰胺，在没有任何共价标记或其他信号传导策略的情况下，已实现了113 nt Capt组装体的高可卡因特异性区分。与控制特性（例如凝胶电泳和荧光共振能量转移（FRET））相比，信噪比得到了大大提高。作为进一步的创新，我们验证了低噪声CGN还可以提高在大尺寸基板侧链上发生的小构象/尺寸变化的分辨率。从杂交链反应（HCR）产生的长双链双键催化剂被选作模型底物。当可卡因存在时，当26 nt适体片段组装在HCR双链体的侧链上时，低噪声CGN可以灵敏地捕获电流变化。

更新日期：2021-03-03

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