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...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/10/2087 |
| _version_ | 1797469594857766912 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Marcel Jaron |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T19:23:33Z |
| format | Article |
| id | doaj.art-059ba81e8dae4b76b6f823fda47e7e79 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T19:23:33Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-059ba81e8dae4b76b6f823fda47e7e792023-11-24T03:07:12ZengMDPI AGViruses1999-49152022-09-011410208710.3390/v14102087Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization AssessmentMarcel Jaron0Michael Lehky1Marta Zarà2Chris Nicole Zaydowicz3Aidin Lak4Rico Ballmann5Philip Alexander Heine6Esther Veronika Wenzel7Kai-Thomas Schneider8Federico Bertoglio9Susanne Kempter10Reinhard Wolfgang Köster11Silvia Stella Barbieri12Joop van den Heuvel13Michael Hust14Stefan Dübel15Maren Schubert16Department of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyRecombinant Protein Expression Platform, Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, GermanyUnit of Brain-Heart Axis, IRCCS Monzino Cardiology Center, Via C. Parea 4, 20138 Milano, ItalyDivision of Cellular and Molecular Neurobiology, Zoological Institute, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyInstitute for Electrical Measurement Science and Fundamental Electrical Engineering, Technische Universität Braunschweig, Hans-Sommer-Straße 66, 38106 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyAbcalis GmbH, Inhoffenstraße 7, 38124 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyDepartment of Physics, Ludwig-Maximilians-Universität, Geschwister-Scholl-Platz 1, 80539 München, GermanyDivision of Cellular and Molecular Neurobiology, Zoological Institute, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyUnit of Brain-Heart Axis, IRCCS Monzino Cardiology Center, Via C. Parea 4, 20138 Milano, ItalyRecombinant Protein Expression Platform, Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyDepartment of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, GermanyVirus-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.https://www.mdpi.com/1999-4915/14/10/2087virus-like particles (VLPs)SARS-CoV-2insect cellsexpression vectorantibodiescellular assay |
| spellingShingle | 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 Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment Viruses virus-like particles (VLPs) SARS-CoV-2 insect cells expression vector antibodies cellular assay |
| title | Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment |
| title_full | Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment |
| title_fullStr | Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment |
| title_full_unstemmed | Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment |
| title_short | Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment |
| title_sort | baculovirus free sars cov 2 virus like particle production in insect cells for rapid neutralization assessment |
| topic | virus-like particles (VLPs) SARS-CoV-2 insect cells expression vector antibodies cellular assay |
| url | https://www.mdpi.com/1999-4915/14/10/2087 |
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