Enhanced Analytes Capture by Engineering Electrostatics at the Entry of Aerolysin Nanopore

Article Article scientifique Article Enhanced Analytes Capture by Engineering Electrostatics at the Entry of Aerolysin Nanopore eng 2025-05-09 2025-05-09 author El Chazli Marwan Institute of Bioengineering School of Life Sciences École Polytechnique Fédérale de Lausanne (EPFL) 1015 Lausanne Switzerland author Bada Juarez Juan Francisco 0000-0002-1337-6424 Institute of Bioengineering School of Life Sciences École Polytechnique Fédérale de Lausanne (EPFL) 1015 Lausanne Switzerland author Perrin Louis Willem 0009-0005-5725-927X Department of Inorganic and Analytical Chemistry Chemistry and Biochemistry University of Geneva 1211 Geneva Switzerland 416484 Chimie minérale groupe 5 (CHAM5) author Gao Jiajie Department of Inorganic and Analytical Chemistry Chemistry and Biochemistry University of Geneva 1211 Geneva Switzerland 408884 author Cao Chan 0000-0003-2592-0690 Department of Inorganic and Analytical Chemistry Chemistry and Biochemistry University of Geneva 1211 Geneva Switzerland 413165 Département de chimie minérale et analytique (CHIAM) e202400180 10.1002/hlca.202400180 0018-019X https://onlinelibrary.wiley.com/doi/10.1002/hlca.202400180 Nanopore technology is a powerful single‐molecule platform for detecting and sequencing a wide range of biomolecules. Among nanopores, aerolysin has emerged as a particularly promising candidate for peptide sensing. However, its ability to capture long biopolymers is limited due to its lack of a vestibule structure. In this study, we engineered electrostatics at the entry of the aerolysin pore and observed an increase in event frequency – up to 2 times higher for DNA and for the peptide compared to wild‐type aerolysin. Importantly, this modification did not affect the pore's current‐voltage characteristics. When tested with DNA and α‐synuclein peptides, the engineered pore (D209R) exhibited comparable dwell times and current blockages to the wild‐type pore, while ion selectivity and electroosmotic flux show an increase. These findings highlight that fine‐tuning the electrostatic properties at the pore entry can significantly enhance event frequency without compromising key transport properties such as current blockage or dwell time. This improvement expands the utility of aerolysin nanopores for sensing and sequencing applications and paves the way for more effective diagnostic tools and analytical methods in the field of proteomics and biomarker discovery. Dewey 540 Swiss National Science Foundation Improving resolution of single-molecule characterization by nanopores 193090 CH Careers 40 Département de chimie minérale, analytique et appliquée 145 false 1445 Groupe Cao Helvetica chimica acta 04296282-f7d7-4d37-a6eb-382327fcda46 Article (Published version) Restricted Helvetica_Chimica_Acta_-_2025_-_El_.pdf 11651368 application/pdf 1398cf1f71e78ed2ab159d89e1913e32c7a24fd3ea0254eb4b61bc3e8266f2f6 restrictedAccess Faculté des sciences / Section de chimie et biochimie / Département de chimie minérale, analytique et appliquée

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Enhanced Analytes Capture by Engineering Electrostatics at the Entry of Aerolysin Nanopore	Helvetica chimica acta			2025	52	1

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