Characterization and applications of a Crimean-Congo hemorrhagic fever virus nucleoprotein-specific Affimer: Inhibitory effects in viral replication and development of colorimetric diagnostic tests.
Álvarez-Rodríguez, Beatriz;
Tiede, Christian;
Hoste, Alexis CR;
Surtees, Rebecca A;
Trinh, Chi H;
Slack, Gillian S;
Chamberlain, John;
Hewson, Roger;
Fresco, Alba;
Sastre, Patricia;
+4 more...Tomlinson, Darren C;
Millner, Paul A;
Edwards, Thomas A;
Barr, John N;
(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 Neglected Tropical Diseases, 14 (6).
e0008364-.
ISSN 1935-2727
DOI: https://doi.org/10.1371/journal.pntd.0008364
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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 Affimer-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 system. 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.