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

Autor: Marcel Jaron, Michael Lehky, Marta Zarà, Chris Nicole Zaydowicz, Aidin Lak, Rico Ballmann, Philip Alexander Heine, Esther Veronika Wenzel, Kai-Thomas Schneider, Federico Bertoglio, Susanne Kempter, Reinhard Wolfgang Köster, Silvia Stella Barbieri, Joop van den Heuvel, Michael Hust, Stefan Dübel, Maren Schubert
Jazyk: angličtina
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Zdroj: Viruses
Viruses; Volume 14; Issue 10; Pages: 2087
Viruses : Special Issue Virus-Like Particle Vaccines 2022 14 (2022) 10, 2087.-https://doi.org/10.3390/v14102087--Viruses--http://www.mdpi.com/journal/viruses--https://www.ncbi.nlm.nih.gov/pmc/journals/1559/--http://www.bibliothek.uni-regensburg.de/ezeit/?2516098--1999-4915--1999-4915
ISSN: 1999-4915
DOI: 10.3390/v14102087
Popis: 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.
Databáze: OpenAIRE
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