Department of Chemical and Materials Engineering, University of Alberta, Edmonton Alberta, Canada.
ACS Appl Bio Mater. 2021 Jul 19;4(7):5416-5424. doi: 10.1021/acsabm.1c00095. Epub 2021 Jun 23.
Hybrid nanopores combine the durability of a solid-state nanopore with the precise structure of a biological nanopore. When a DNA strand is pulled electrophoretically through a solid-state nanopore it can be sensed using the ionic blockade current produced by each translocating molecule. However, owing to the lack of chemical specificity and pore size reproducibility, solid-state nanopore sensing suffers from poor repeatability. Biological nanopores which have a constant geometry ensure sensitive and repeatable sensing. In this study, hybrid nanopores were formed by insertion of a engineered outer membrane porin G (eOmpG) in a bilayer (BL) molybdenum disulfide (MoS) solid-state nanopore. Engineered outer membrane porin G (eOmpG) is used as the biological counterpart of the hybrid nanopore due to its uniform cylindrical geometry and controlled gating useful for specific detection of label-free analytes. BL MoS is used as the solid-state support for the hybrid construct owing to its surface charge and 2D layered properties, which ensures a stable support with low capacitive noise, favorable for precise sensing. To realize the hybried nanopore a single eOmpG was electrophoretically pulled through a 3.4 nm BL MoS solid-state nanopore at neutral pH and +80 mV trans bias. A hybrid BL MoS-eOmpG nanopore was found to demonstrate 32% lower noise levels with nearly 1.9 times improved in the signal-to-noise ratio (SNR) and 6.5 times longer dwell times for dA30 molecular sensing compared to the BL MoS solid-state nanopore. Thus, the low-noise biocompatible platform of the hybrid BL MoS-eOmpG nanopore can be used for highly resolved biomolecular sensing.
杂化纳米孔结合了固态纳米孔的耐用性和生物纳米孔的精确结构。当 DNA 链通过固态纳米孔电泳拉伸时,可以使用每个迁移分子产生的离子阻塞电流来检测它。然而,由于缺乏化学特异性和孔尺寸重现性,固态纳米孔传感的重复性较差。具有恒定几何形状的生物纳米孔确保了敏感和可重复的传感。在这项研究中,通过将工程化的外膜孔蛋白 G(eOmpG)插入双层(BL)二硫化钼(MoS)固态纳米孔中形成杂化纳米孔。工程化的外膜孔蛋白 G(eOmpG)用作杂化纳米孔的生物对应物,因为其具有均匀的圆柱形几何形状和受控的门控,可用于对无标记分析物进行特异性检测。BL MoS 用作杂化结构的固态支撑物,因为其表面电荷和 2D 层状特性可确保具有低电容噪声的稳定支撑,有利于精确传感。为了实现杂化纳米孔,在中性 pH 值和+80 mV 跨偏压下,通过 3.4nm BL MoS 固态纳米孔电泳拉制单个 eOmpG。与 BL MoS 固态纳米孔相比,杂化 BL MoS-eOmpG 纳米孔的噪声水平降低了 32%,信号噪声比(SNR)提高了近 1.9 倍,dA30 分子传感的停留时间延长了 6.5 倍。因此,杂化 BL MoS-eOmpG 纳米孔的低噪声生物相容性平台可用于高分辨率生物分子传感。