Di Genova, Cecilia, Sampson, Alex, Scott, Simon, Cantoni, Diego, Mayora-Neto, Martin, Bentley, Emma, Mattiuzzo, Giada, Wright, Edward, Derveni, Mariliza, Auld, Bethany, Harrison, Dale, Ferrara, Bill T. ORCID: https://orcid.org/0000-0002-2163-4032, Said, Mohamed, Selim, Arwa ORCID: https://orcid.org/0000-0001-5552-7561, Thompson, Elinor ORCID: https://orcid.org/0000-0002-6434-9290, Thompson, Craig, Carnell, George and Temperton, Nigel ORCID: https://orcid.org/0000-0002-7978-3815 (2021) Production, titration, neutralisation and storage of SARS-CoV-2 lentiviral pseudotypes. Bio-protocol, 11 (21). pp. 1-21. ISSN 2331-8325 (doi:10.21769/BioProtoc.4236)

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Abstract

This protocol details a rapid and reliable method for the production and titration of high-titer viral pseudotype particles with the SARS-CoV-2 spike protein (and D614G variant) on a lentiviral vector core, and use for neutralization assays in target cells expressing ACE2 and TMPRSS2. It additionally provides detailed instruction on substituting in new spike variants via gene cloning, and storage/shipping considerations for wide deployment potential. Results obtained with this protocol show that SARS-CoV-2 pseudotypes can be produced at equivalent titers to SARS-CoV and MERS-CoV pseudotypes, that they can be neutralized by human convalescent plasma and monoclonal antibodies and that they can be stored at a range of laboratory temperatures and lyophilized for distribution.

Item Type:	Article
Additional Information:	Bio-protocol articles deriving from eLife papers are distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use and redistribution provided that the original author and source are credited.
Uncontrolled Keywords:	Covid-19; titration; neutralisation; storage; SARS-CoV-2; pseudotype; antibody
Subjects:	Q Science > QC Physics Q Science > QD Chemistry
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science Faculty of Engineering & Science > Medway School of Pharmacy Faculty of Engineering & Science > School of Science (SCI)
Related URLs:	Publisher
Last Modified:	11 May 2023 14:38
URI:	http://gala.gre.ac.uk/id/eprint/30820

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