Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment

Virus-like particles (VLPs) resemble authentic virus while not containing any genomic information. Here, we present a fast and powerful method for the production of SARS-CoV-2 VLP in insect cells and the application of these VLPs to evaluate the inhibition capacity of monoclonal antibodies and sera...

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Bibliographic Details
Published in:	Viruses Vol. 14; no. 10; p. 2087
Main Authors:	Jaron, Marcel, Lehky, Michael, Zarà, Marta, Zaydowicz, Chris Nicole, Lak, Aidin, Ballmann, Rico, Heine, Philip Alexander, Wenzel, Esther Veronika, Schneider, Kai-Thomas, Bertoglio, Federico, Kempter, Susanne, Köster, Reinhard Wolfgang, Barbieri, Silvia Stella, van den Heuvel, Joop, Hust, Michael, Dübel, Stefan, Schubert, Maren
Format:	Journal Article
Language:	English
Published:	Basel MDPI AG 20-09-2022 MDPI
Subjects:	ACE2 Angiotensin Angiotensin II Angiotensin II receptor blockers Angiotensin-converting enzyme 2 antibodies Baculoviruses cellular assay Coronaviruses COVID-19 expression vector Flow cytometry Health aspects Insect cells Laboratories Membrane fusion Membrane proteins Microscopy Molecular weight Monoclonal antibodies Nanoparticles Peptides Plasmids Prevention Proteins SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 Spike protein Sucrose Transfection Transmission electron microscopy Vaccines Virus-like particles virus-like particles (VLPs) Viruses Germany United States > US
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Summary:	Virus-like particles (VLPs) resemble authentic virus while not containing any genomic information. Here, we present a fast and powerful method for the production of SARS-CoV-2 VLP in insect cells and the application of these VLPs to evaluate the inhibition capacity of monoclonal antibodies and sera of vaccinated donors. Our method avoids the baculovirus-based approaches commonly used in insect cells by employing direct plasmid transfection to co-express SARS-CoV-2 envelope, membrane, and spike protein that self-assemble into VLPs. After optimization of the expression plasmids and vector ratios, VLPs with an ~145 nm diameter and the typical “Corona” aura were obtained, as confirmed by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Fusion of the membrane protein to GFP allowed direct quantification of binding inhibition to angiotensin II-converting enzyme 2 (ACE2) on cells by therapeutic antibody candidates or sera from vaccinated individuals. Neither VLP purification nor fluorescent labeling by secondary antibodies are required to perform these flow cytometric assays.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1999-4915 1999-4915
DOI:	10.3390/v14102087