Genova, Cecilia, Sampson, Alex, Scott, Simon, Cantoni, Diego, Neto, Martin, Bentley, Emma, Mattiuzzo, Giada, Wright, Edward, Derveni, Mariliza, Auld, Bethany, and others. (2021) Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes. BIO-PROTOCOL, 11 (21). ISSN 2331-8325. (doi:10.21769/BioProtoc.4236) (KAR id:91352)
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Official URL https://doi.org/10.21769/BioProtoc.4236 |
Abstract
This protocol details a rapid and reliable method for the production and titration of high-titre viral pseudotype particles with the SARS-CoV-2 spike protein (and D614G or other variants of concern, VOC) on a lentiviral vector core, and use for neutralisation assays in target cells expressing angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2). It additionally provides detailed instructions on substituting in new spike variants via gene cloning, lyophilisation and storage/shipping considerations for wide deployment potential. Results obtained with this protocol show that SARS-CoV-2 pseudotypes can be produced at equivalent titres to SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV) pseudotypes, neutralised by human convalescent plasma and monoclonal antibodies, and stored at a range of laboratory temperatures and lyophilised for distribution and subsequent application.
Item Type: | Article |
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DOI/Identification number: | 10.21769/BioProtoc.4236 |
Subjects: | Q Science > QR Microbiology > QR355 Virology |
Divisions: | Divisions > Division of Natural Sciences > Medway School of Pharmacy |
Depositing User: | Nigel Temperton |
Date Deposited: | 05 Nov 2021 09:47 UTC |
Last Modified: | 17 Dec 2021 23:10 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/91352 (The current URI for this page, for reference purposes) |
Temperton, Nigel J.: | ![]() |
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