Nanopore-Based Devices for Bioanalytical Applications

Abstract

W ith over a decade passed since the first reported use of a Staphylococcal a-hemolysin pore to study single molecules of single-stranded DNA, research in the field of nanopores has advanced rapidly. We discuss the technological progression of nanopore-based devices from the initial use of a-hemolysin pores to the advent of solid-state nanopores to the burgeoning of organice inorganic hybrid pores driven by the desire to achieve fast and inexpensive DNA sequencing. Additional nanoporebased efforts are also discussed that study other classes of molecules, such as proteins. We discuss the use of nanopores for protein folding and binding analysis. In addition to single-molecule analysis, we report on the introduction of nanopore arrays on thin film membranes for ultrafiltration. Owing to their reduced spatial dimensionality, such membranes offer greater control over how the pores interact with analytes thus leading to very efficient separation. With several technical hindrances yet to be overcome, the devices we report are still works in progress. The realization of these devices will enhance laboratory processes by permitting superior spatial and temporal analytical resolution at the single-molecule level resulting in laboratory capacities of great impact.

Layer-by-layer removal of insulating few-layer mica flakes for asymmetric ultra-thin nanopore fabrication

“…In particular, in the applications of nanopore-based analysis and separation [15], ultra-thin nanopores with both length and diameter matching the size of molecular analytes show promising superiorities over previous reported nanopore systems [16][17][18][19][20][21][22]. Although the diameter of the state-of-the-art SiN nanopores can be successfully reduced down to 1.5-2 nm, a substantial challenge remains in that none of them has a channel length shorter than 5 nm [23][24][25][26].…”

Amplification of Light-induced Molecular-Shape Change by Supramolecular Machines

“…The pore is inherently stable and acts as a nonspecific ion channel for application to some engineered tools. 8,9 We have already prepared a chimeric protein N-PYP-Hla, where PYP is connected to the N-termini of Hla, and found that the hemolytic activity of this chimeric protein could be controlled by visible-light irradaition. 10 Here we report construction of chimeric protein C-PYP-Hla, where PYP is connected to the C-termini of Hla, and its hemolytic activity in the dark and under irradiation.…”

Emerging trends in precision fabrication of microapertures to support suspended lipid membranes for sensors, sequencing, and beyond

“…Due to the natural distribution of ion channels, and the corresponding ligand diversity, ion channels have been explored as sensors for hormones, neurotransmitters and other natural and synthetic targets [5–7]. Additionally, ion channels play a key role in the development of next generation single molecule DNA sequencing [8–10]. …”

“…BLMs span an aperture that connects two otherwise isolated fluidic compartments (Figure 1), allowing fluidic and electronic access to both faces of the membrane, in a manner similar to traditional cellular patch clamp methodologies. The largest success of BLMs, to date, is the demonstrated utility in ion channel-based nanopore DNA sequencing [8–10]. Often, large diameter apertures, and thus large area BLMs, have been utilized, though smaller diameter apertures have been demonstrated to improve bilayer performance [13, 14].…”