Solid-state nanopore based biomimetic voltage gated ion channels.

Voltage gating is essential to the computational ability of neurons. We show this effect can be mimicked in a solid-state nanopore by functionalizing the pore interior with a redox active molecule. We study the integration of an active biological molecule-a quinone-into a solid state nanopore, and its subsequent induced voltage gating. We show that the voltage gating effect mimics biological gating systems in its classic sigmoidal voltage response, unlike previous synthetic voltage gating systems. Initially, the quinone undergoes a reduction due to radicals in the bulk solution, and is converted to the hydroquinone state. Upon deprontonation the hydroquinone then acts as a charged nanomechanical arm, which opens the channel under the applied potential. We establish that the quinone gains a single net charge when the pH inside of the nanopore reaches its pKa value, and explore factors that influence the net pH in the middle of the pore. Using a combination of theory, experiment and simulation, we conclude that concentration polarization and a shift of the pH inside of the channel is the main cause of this gating effect.

Tailored Polymeric Membranes for Mycobacterium Smegmatis Porin A (MspA) Based Biosensors.

Nanopores based on protein channels inserted into lipid membranes have paved the way towards a wide-range of inexpensive biosensors, especially for DNA sequencing. A key obstacle in using these biological ion channels as nanodevices is the poor stability of lipid bilayer membranes. Amphiphilic block copolymer membranes have emerged as a robust alternative to lipid membranes. While previous efforts have shown feasibility, we demonstrate for the first time the effect of polymer composition on MspA protein functionality. We show that membrane-protein interaction depends on the hydrophobic-hydrophilic ratio (f-ratio) of the block copolymer. These effects are particularly pronounced in asymmetric protein pores like MspA compared to the cylindrical α-Hemolysin pore. A key effect of membrane-protein interaction is the increased 1/fα noise. After first showing increases in 1/fα behaviour arise from increased substate activity, the noise power spectral density S(f) was used as a qualitative tool for understanding protein-membrane interactions in polymer membranes. Polymer compositions with f-ratios close to lipid membranes caused noise behaviour not observed in lipid membranes. However, by modifying the f-ratio using a modular synthetic approach, we were able to design a block copolymer exhibiting noise properties similar to a lipid membrane, albeit with better stability. Thus, by careful optimization, block copolymer membranes can emerge as a robust alternative for protein-pore based nano-biosensors.

A CMOS enhanced solid-state nanopore based single molecule detection platform.

Solid-state nanopores have emerged as a single molecule label-free electronic detection platform. Existing transimpedance stages used to measure ionic current nanopores suffer from dynamic range limitations resulting from steady-state baseline currents. We propose a digitally-assisted baseline cancellation CMOS platform that circumvents this issue. Since baseline cancellation is a form of auto-zeroing, the 1/f noise of the system is also reduced. Our proposed design can tolerate a steady state baseline current of 10µA and has a usable bandwidth of 750kHz. Quantitative DNA translocation experiments on 5kbp DNA was performed using a 5nm silicon nitride pore using both the CMOS platform and a commercial system. Comparison of event-count histograms show that the CMOS platform clearly outperforms the commercial system, allowing for unambiguous interpretation of the data.