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Nano Research 2022, 15(11): 9936-9942 https://doi.org/10.1007/s12274-022-4631-9
Research Article | Issue | Published: 04 July 2022

Effects of symmetric and asymmetric salt conditions on a selective solid-state nanopore assay

Show Author's Information Ian D. Wadsworth1,  Adam R. Hall1,2( )
Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA
Comprehensive Cancer Center, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA
Keywords:
biomarkers, solid-state nanopores, nucleic acids, amplification-free detection, salt gradient
Topics:
Nanopores
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Wadsworth ID, Hall AR. Effects of symmetric and asymmetric salt conditions on a selective solid-state nanopore assay. Nano Research, 2022, 15(11): 9936-9942. https://doi.org/10.1007/s12274-022-4631-9
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Abstract Full text Electronic supplementary material About this article

Conventional solid-state nanopore measurements sense all translocating entities, necessitating meticulous analysis to differentiate target biomolecules. To address this, we have established a selective assay with the platform that has shown utility in quantifying several nucleic acid biomarkers. However, limited detection efficiency and intrinsic noise have so far limited assay resolution to 10 nM. Improvements in this value require manipulation of translocation dynamics. Here, we report the effects of NaCl conditions on assay performance. We first investigate symmetric conditions, finding sensitivity increases with salt concentration but selectivity is maximized at 1.0 M NaCl. We then probe asymmetric conditions, showing a remarkable impact on assay sensitivity and selectivity when measurement buffer NaCl concentration in the reservoir with the translocating molecules is low and the opposite reservoir is increased. Using optimum conditions, we demonstrate detection of target biomolecules down to a concentration of 100 pM which is an improvement of 2 orders of magnitude over past results.


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Electronic supplementary material
About this article

Effects of symmetric and asymmetric salt conditions on a selective solid-state nanopore assay

Show Author's information Ian D. Wadsworth1, Adam R. Hall1,2( )
Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA
Comprehensive Cancer Center, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA

Abstract

Conventional solid-state nanopore measurements sense all translocating entities, necessitating meticulous analysis to differentiate target biomolecules. To address this, we have established a selective assay with the platform that has shown utility in quantifying several nucleic acid biomarkers. However, limited detection efficiency and intrinsic noise have so far limited assay resolution to 10 nM. Improvements in this value require manipulation of translocation dynamics. Here, we report the effects of NaCl conditions on assay performance. We first investigate symmetric conditions, finding sensitivity increases with salt concentration but selectivity is maximized at 1.0 M NaCl. We then probe asymmetric conditions, showing a remarkable impact on assay sensitivity and selectivity when measurement buffer NaCl concentration in the reservoir with the translocating molecules is low and the opposite reservoir is increased. Using optimum conditions, we demonstrate detection of target biomolecules down to a concentration of 100 pM which is an improvement of 2 orders of magnitude over past results.

Keywords: biomarkers, solid-state nanopores, nucleic acids, amplification-free detection, salt gradient

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Publication history
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Acknowledgements

Publication history

Received: 20 December 2021
Revised: 09 May 2022
Accepted: 31 May 2022
Published: 04 July 2022
Issue date: November 2022

Copyright

© Tsinghua University Press 2022

Acknowledgements

Acknowledgements

This project was supported by NIH awards (Nos. R21CA193067, R33CA246448, and P41EB020594). SS-nanopore fabrication was performed at the Rutgers University Laboratory for Surface Modification. We acknowledge the laboratory of Dr. Mark Howarth (Oxford University) for providing MS protein. We thank Mallory Smith for contributions to figures.

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