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Peptides. 2017 Feb;88:87-96. doi: 10.1016/j.peptides.2016.12.015. Epub 2016 Dec 22.

An enzymatic assay based on luciferase Ebola virus-like particles for evaluation of virolytic activity of antimicrobial peptides.

Author information

1
Central European Institute of Technology (CEITEC), Masaryk University, Kamenice 5, 625 00 Brno, Czechia. Electronic address: m.peskova@mail.muni.cz.
2
Department of Chemistry and Biochemistry, Mendel University, 613 00 Brno, Czechia; Central European Institute of Technology (CEITEC), Brno University of Technology, 616 00 Brno, Czechia. Electronic address: zbynek.heger@mendelu.cz.
3
Central European Institute of Technology (CEITEC), Masaryk University, Kamenice 5, 625 00 Brno, Czechia. Electronic address: 323853@mail.muni.cz.
4
Department of Chemistry and Biochemistry, Mendel University, 613 00 Brno, Czechia; Central European Institute of Technology (CEITEC), Brno University of Technology, 616 00 Brno, Czechia. Electronic address: vojtech.adam@mendelu.cz.
5
Central European Institute of Technology (CEITEC), Masaryk University, Kamenice 5, 625 00 Brno, Czechia; Institute of Physiology, Faculty of Medicine, Masaryk University, Kamenice 5, 625 00 Brno, Czechia. Electronic address: pekarikv@mail.muni.cz.

Abstract

Antimicrobial peptides are currently considered as promising antiviral compounds. Current assays to evaluate the effectivity of peptides against enveloped viruses based on liposomes or hemolysis are encumbered by the artificial nature of liposomes or distinctive membrane composition of used erythrocytes. We propose a novel assay system based on enzymatic Ebola virus-like particles containing sensitive luciferase reporter. The assay was validated with several cationic and anionic peptides and compared with lentivirus inactivation and hemolytic assays. The assay is sensitive and easy to perform in standard biosafety level laboratory with potential for high-throughput screens. The use of virus-like particles in the assay provides a system as closely related to the native viruses as possible eliminating some issues associated with other more artificial set ups. We have identified CAM-W (KWKLWKKIEKWGQGIGAVLKWLTTWL) as a peptide with the greatest antiviral activity against infectious lentiviral vectors and filoviral virus-like particles.

KEYWORDS:

Antimicrobial peptides; Cytotoxic peptides; Ebola; Hemolytic activity; Lentivirus; Marburg; Nanoluciferase; Virolytic activity; Virus-like particles

PMID:
28012857
DOI:
10.1016/j.peptides.2016.12.015
[Indexed for MEDLINE]

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