Chau, CC orcid.org/0000-0002-3134-6798, Radford, SE orcid.org/0000-0002-3079-8039, Hewitt, EW orcid.org/0000-0002-6238-6303 et al. (1 more author) (2020) Macromolecular Crowding Enhances the Detection of DNA and Proteins by a Solid-State Nanopore. Nano Letters, 20 (7). pp. 5553-5561. ISSN 1530-6984
Abstract
Nanopore analysis of nucleic acid is now routine, but detection of proteins remains challenging. Here, we report the systematic characterization of the effect of macromolecular crowding on the detection sensitivity of a solid-state nanopore for circular and linearized DNA plasmids, globular proteins (β-galactosidase), and filamentous proteins (α-synuclein amyloid fibrils). We observe a remarkable ca. 1000-fold increase in the molecule count for the globular protein β-galactosidase and a 6-fold increase in peak amplitude for plasmid DNA under crowded conditions. We also demonstrate that macromolecular crowding facilitates the study of the topology of DNA plasmids and the characterization of amyloid fibril preparations with different length distributions. A remarkable feature of this method is its ease of use; it simply requires the addition of a macromolecular crowding agent to the electrolyte. We therefore envision that macromolecular crowding can be applied to many applications in the analysis of biomolecules by solid-state nanopores.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2020 American Chemical Society. This is an open access article under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) (https://creativecommons.org/licenses/by/4.0/) |
Keywords: | Nanopore, nanopipette, macromolecular crowding, single-molecule, DNA |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Electronic & Electrical Engineering (Leeds) > Pollard Institute (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology (Leeds) |
Funding Information: | Funder Grant number EU - European Union 322408 Royal Society RG160273 EPSRC (Engineering and Physical Sciences Research Council) EP/S01764X/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 24 Jun 2020 12:51 |
Last Modified: | 20 Dec 2024 14:38 |
Status: | Published |
Publisher: | American Chemical Society |
Identification Number: | 10.1021/acs.nanolett.0c02246 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:162126 |