Characterization and applications of a crimean-congo hemorrhagic fever virus nucleoprotein-specific affimer: Inhibitory effects in viral replication and development of colorimetric diagnostic tests

Beatriz Álvarez-Rodríguez, Christian Tiede, Alexis C.R. Hoste, Rebecca A. Surtees, Chi H. Trinh, Gillian S. Slack, John Chamberlain, Roger Hewson, Alba Fresco, Patricia Sastre, Darren C. Tomlinson, Paul A. Millner, Thomas A. Edwards*, John N. Barr

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)
7 Downloads (Pure)

Abstract

Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV) is one of the most widespread medically important arboviruses, causing human infections that result in mortality rates of up to 60%. We describe the selection of a high-affinity small protein (Affimer-NP) that binds specifically to the nucleoprotein (NP) of CCHFV. We demonstrate the interference of Affi-mer-NP in the RNA-binding function of CCHFV NP using fluorescence anisotropy, and its inhibitory effects on CCHFV gene expression in mammalian cells using a mini-genome sys-tem. Solution of the crystallographic structure of the complex formed by these two molecules at 2.84 Å resolution revealed the structural basis for this interference, with the Affimer-NP binding site positioned at the critical NP oligomerization interface. Finally, we validate the in vitro application of Affimer-NP for the development of enzyme-linked immunosorbent and lateral flow assays, presenting the first published point-of-care format test able to detect recombinant CCHFV NP in spiked human and animal sera.

Original languageEnglish
Article numbere0008364
Pages (from-to)1-20
Number of pages20
JournalPLoS Neglected Tropical Diseases
Volume14
Issue number6
DOIs
Publication statusPublished - 3 Jun 2020

Bibliographical note

Funding Information: This work was supported by an EU Marie Skłodowska-Curie Actions (MSCA, ec.europa.eu) Innovative Training Network (ITN): H2020- MSCA-ITN-2016, under grant No 721367 (to B. Álvarez-Rodríguez). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Open Access: This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Publisher Copyright: © 2020 Álvarez-Rodríguez et al.

Citation: Álvarez-Rodríguez B, Tiede C, Hoste ACR, Surtees RA, Trinh CH, Slack GS, et al. (2020) Characterization and applications of a Crimean-Congo hemorrhagic fever virus nucleoprotein-specific Affimer: Inhibitory effects in viral replication and development of colorimetric diagnostic tests. PLoS Negl Trop Dis 14(6): e0008364.

DOI: https://doi.org/10.1371/journal.pntd.0008364

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