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Characterization and applications of a Crimean-Congo hemorrhagic fever virus nucleoprotein-specific Affimer: Inhibitory effects in viral replication and development of colorimetric diagnostic tests

DOI: 10.1371/journal.pntd.0008364 DOI Help

Authors: Beatriz Alvarez-Rodriguez (University of Leeds) , Christian Tiede (University of Leeds) , Alexis C. R. Hoste (University of Leeds) , Rebecca A. Surtees (University of Leeds) , Chi H. Trinh (University of Leeds) , Gillian S. Slack (Public Health England) , John Chamberlain (Public Health England) , Roger Hewson (Public Health England) , Alba Fresco (INGENASA, Inmunología y Genética Aplicada S.A.) , Patricia Sastre (INGENASA, Inmunología y Genética Aplicada S.A.) , Darren C. Tomlinson (University of Leeds) , Paul A. Millner (University of Leeds) , Thomas A. Edwards (University of Leeds) , John N. Barr (University of Leeds)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Plos Neglected Tropical Diseases , VOL 14

State: Published (Approved)
Published: June 2020

Open Access Open Access

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 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.

Journal Keywords: Crystal structure; Enzyme-linked immunoassays; RNA structure; Gene expression; Latex; Binding analysis; Genetic interference; Molecular structure

Diamond Keywords: Crimean-Congo Hemorrhagic Fever (CCHF); Viruses

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I03-Macromolecular Crystallography

Documents:
journal.pntd.0008364.pdf

Discipline Tags:

Pathogens Infectious Diseases Disease in the Developing World Health & Wellbeing Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)