Single Molecule Nanopore Spectrometry for Peptide Detection

Published

August 15, 2017

Author(s)

Amy Chavis, Joseph Robertson, Joseph E. Reiner, Kyle Brady, Grrace Hatmaker, Nuwan Kothalawala

Abstract

Sensing and characterization of water-soluble peptides is of critical importance in a wide variety of bioapplications. Single molecule nanopore spectrometry (SMNS) is based on the idea that one can use biological protein nanopores to resolve different sized molecules down to limits set by the blockade duration and noise. Previous work has shown that this enables discrimination between polyethylene glycol (PEG) molecules that differ by a single monomer unit. This paper describes efforts to extend SMNS to a variety of biologically relevant, water-soluble peptides. We describe the use of Au25(SG)18 clusters, previously shown to improve PEG detection, to increase the on- and off-rate of peptides to the pore. In addition, we study the role that fluctuations play in the single molecule nanopore spectrometry (SMNS) methodology and show that modifying solution conditions to increase peptide flexibility (via pH or chaotropic salt) leads to a nearly 2-fold reduction in the current blockade fluctuations and a corresponding narrowing of the peaks in the blockade distributions. Finally, a model is presented that connects the current blockade depths to the mass of the peptides, which shows that our enhanced SMNS detection improves the mass resolution of the nanopore sensor more than 2-fold for the largest cationic peptides studied.

Citation

ACS Sensors

Pub Type

Journals

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DOI Link

Citation

Chavis, A. , Robertson, J. , Reiner, J. , Brady, K. , Hatmaker, G. and Kothalawala, N. (2017), Single Molecule Nanopore Spectrometry for Peptide Detection, ACS Sensors, [online], https://doi.org/10.1021/acssensors.7b00362 (Accessed May 10, 2025)