Development of a NS2B/NS3 protease inhibition assay using AlphaScreen® beads for screening of anti-dengue activities

Muhammad Asyraf Abduraman; Maywan Hariono; Rohana Yusof; Noorsaadah Abd Rahman; Habibah A. Wahab; Mei Lan Tan

Heliyon (Dec 2018)

Development of a NS2B/NS3 protease inhibition assay using AlphaScreen® beads for screening of anti-dengue activities

Muhammad Asyraf Abduraman,
Maywan Hariono,
Rohana Yusof,
Noorsaadah Abd Rahman,
Habibah A. Wahab,
Mei Lan Tan

Affiliations

Muhammad Asyraf Abduraman: Malaysian Institute of Pharmaceuticals & Nutraceuticals, National Institutes of Biotechnology Malaysia (NIBM), Ministry of Science, Technology and Innovation Malaysia, Pulau Pinang, Malaysia; Advanced Medical & Dental Institute, Universiti Sains Malaysia, Pulau Pinang, Malaysia
Maywan Hariono: Pharmaceuticals Drug Simulation Laboratory (PhDS), School of Pharmaceutical Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia
Rohana Yusof: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
Noorsaadah Abd Rahman: Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
Habibah A. Wahab: Malaysian Institute of Pharmaceuticals & Nutraceuticals, National Institutes of Biotechnology Malaysia (NIBM), Ministry of Science, Technology and Innovation Malaysia, Pulau Pinang, Malaysia; Pharmaceuticals Drug Simulation Laboratory (PhDS), School of Pharmaceutical Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia; Corresponding author.
Mei Lan Tan: Malaysian Institute of Pharmaceuticals & Nutraceuticals, National Institutes of Biotechnology Malaysia (NIBM), Ministry of Science, Technology and Innovation Malaysia, Pulau Pinang, Malaysia; Advanced Medical & Dental Institute, Universiti Sains Malaysia, Pulau Pinang, Malaysia; Corresponding author.

Journal volume & issue: Vol. 4, no. 12
p. e01023

Abstract

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Background: Dengue infection is an endemic infectious disease and it can lead to dengue fever, dengue hemorrhagic fever, and/or dengue shock syndromes. Dengue NS2B/NS3 protease complex is essential for viral replication and is a primary target for anti-dengue drug development. In this study, a NS2B/NS3 protease inhibition assay was developed using AlphaScreen® beads and was used to screen compounds for their protease inhibition activities. Methods: The assay system utilized a known NS2B/NS3 peptide substrate, a recombinant of NS2B/NS3 protease with proprietary StrepTactin® donor and nickel chelate acceptor beads in 384-well format. Results: The optimized assay to screen for NS2B/NS3 protease inhibitors was demonstrated to be potentially useful with reasonable zʹ factor, coefficient variance and signal to background ratio. However, screening of synthesized thioguanine derivatives using the optimized AlphaScreen® assay revealed weak NS2B/NS3 inhibition activities. Conclusion: The AlphaScreen® assay to screen for NS2B/NS3 protease inhibitors is potentially applicable for high throughput screening.

Published in Heliyon

ISSN: 2405-8440 (Online)
Publisher: Elsevier
Country of publisher: United Kingdom
LCC subjects: Science: Science (General); Social Sciences: Social sciences (General)
Website: https://www.cell.com/heliyon/home

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