Nahas, K A; Fletcher, M; Hammond, K; Nehls, C; Cama, J; Ryadnov, M G; Keyser, U F (2022) Measuring thousands of single vesicle leakage events reveals the mode of action of antimicrobial peptides. Analytical Chemistry, 94 (27). pp. 9530-9539.

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Abstract

Host defence or antimicrobial peptides hold promise for providing new pipelines of effective antimicrobial agents. Their activity quantified against reconstituted phospholipid membranes is fundamental to a detailed understanding of their structure-activity relationships. However, existing characterisation assays lack the resolution necessary to achieve this insight. By leveraging a high-throughput and highly parallelized microfluidic platform for trapping and studying thousands of giant unilamellar vesicles, we conducted quantitative long-term microscopy studies to monitor the membrane-disruptive activity of archetypal antimicrobial peptides and bacteriocins with a high spatiotemporal resolution. We correlated the modes of action of 11 distinct peptides with the behaviour of disrupted vesicle populations under the conditions of continuous peptide dosing using a range of concentrations. We further related the observed modes of action to the molecular activity mechanisms of these peptides observed by atomic force microscopy. The study offers an effective approach for screening antimicrobial agents versus the modes of their action.

Item Type:	Article
Subjects:	Biotechnology > Biopharmaceutical Manufacturing and Characterisation
Divisions:	Chemical & Biological Sciences
Identification number/DOI:	10.1021/acs.analchem.1c03564
Last Modified:	12 Sep 2022 13:46
URI:	https://eprintspublications.npl.co.uk/id/eprint/9513