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

doi:10.1016/j.heliyon.2018.e01023

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

Heliyon. 2018 Dec 8;4(12):e01023. doi: 10.1016/j.heliyon.2018.e01023. eCollection 2018 Dec.

Authors

Muhammad Asyraf Abduraman^{1

2}, Maywan Hariono³, Rohana Yusof⁴, Noorsaadah Abd Rahman⁵, Habibah A Wahab^{1

3}, Mei Lan Tan^{1

2}

Affiliations

¹ Malaysian Institute of Pharmaceuticals & Nutraceuticals, National Institutes of Biotechnology Malaysia (NIBM), Ministry of Science, Technology and Innovation Malaysia, Pulau Pinang, Malaysia.
² Advanced Medical & Dental Institute, Universiti Sains Malaysia, Pulau Pinang, Malaysia.
³ Pharmaceuticals Drug Simulation Laboratory (PhDS), School of Pharmaceutical Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia.
⁴ Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
⁵ Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia.

Abstract

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 z' 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.

Keywords: Bioinformatics; Biotechnology.