Direct and Ultrasensitive Bioluminescent Detection of Intact Respiratory Viruses

Alexander Gräwe; Harm van der Veer; Seino A.K. Jongkees; Jacky Flipse; Iebe Rossey; Robert P. de Vries; Xavier Saelens; Maarten Merkx

doi:10.1021/acssensors.4c01855

Direct and Ultrasensitive Bioluminescent Detection of Intact Respiratory Viruses

Alexander Gräwe, Harm van der Veer, Seino A.K. Jongkees, Jacky Flipse, Iebe Rossey, Robert P. de Vries, Xavier Saelens, Maarten Merkx^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Respiratory viruses such as SARS-CoV-2, influenza, and respiratory syncytial virus (RSV) represent pressing health risks. Rapid diagnostic tests for these viruses detect single antigens or nucleic acids, which do not necessarily correlate with the amount of the intact virus. Instead, specific detection of intact respiratory virus particles may be more effective at assessing the contagiousness of a patient. Here, we report GLOVID, a modular biosensor platform to detect intact virions against a background of “free” viral proteins in solution. Our approach harnesses the multivalent display of distinct proteins on the surface of a viral particle to template the reconstitution of a split luciferase, allowing specific, single-step detection of intact influenza A and RSV virions corresponding to 0.1-0.3 fM of genomic units. The protein ligation system used to assemble GLOVID sensors is compatible with a broad range of binding domains, including nanobodies, scFv fragments, and cyclic peptides, which allows straightforward adjustment of the sensor platform to target different viruses.

Original language	English
Pages (from-to)	5550-5560
Number of pages	11
Journal	ACS Sensors
Volume	9
Issue number	10
Early online date	27 Sept 2024
DOIs	https://doi.org/10.1021/acssensors.4c01855
Publication status	Published - 25 Oct 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors. Published by American Chemical Society.

Funding

The authors thank the group of Bruno Correia for providing purified, prefusion-stabilized RSV-F and thank Pramila Rijal and Alain Townsend for kindly providing N1/09 and the sequence of the N2-specific AS4C antibody. We would also like to thank the Research Group Applied Natural Sciences of Fontys University of Applied Sciences Eindhoven for access to ddPCR and the members of the Merkx group for fruitful discussions and feedback. This project has received funding from the European Union\u2019s Horizon 2020 research and innovation program under the Marie Sk\u0142odowska-Curie Grant Agreement No. 899987. This project has received funding from the European Union\u2019s Horizon 2020 research and innovation program under the Marie Sk\u0142odowska-Curie Grant Agreement No. 899987.

Funders	Funder number
Horizon 2020 Framework Programme
H2020 Marie Skłodowska-Curie Actions	899987
H2020 Marie Skłodowska-Curie Actions

Keywords

bioluminescence
biosensor
protein engineering
protein ligation
Respiratory virus
virus diagnostics

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title = "Direct and Ultrasensitive Bioluminescent Detection of Intact Respiratory Viruses",

abstract = "Respiratory viruses such as SARS-CoV-2, influenza, and respiratory syncytial virus (RSV) represent pressing health risks. Rapid diagnostic tests for these viruses detect single antigens or nucleic acids, which do not necessarily correlate with the amount of the intact virus. Instead, specific detection of intact respiratory virus particles may be more effective at assessing the contagiousness of a patient. Here, we report GLOVID, a modular biosensor platform to detect intact virions against a background of “free” viral proteins in solution. Our approach harnesses the multivalent display of distinct proteins on the surface of a viral particle to template the reconstitution of a split luciferase, allowing specific, single-step detection of intact influenza A and RSV virions corresponding to 0.1-0.3 fM of genomic units. The protein ligation system used to assemble GLOVID sensors is compatible with a broad range of binding domains, including nanobodies, scFv fragments, and cyclic peptides, which allows straightforward adjustment of the sensor platform to target different viruses.",

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AU - Rossey, Iebe

AU - de Vries, Robert P.

AU - Saelens, Xavier

AU - Merkx, Maarten

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Direct and Ultrasensitive Bioluminescent Detection of Intact Respiratory Viruses

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Keywords

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