Journal article
Bio-protocol, 2021
Chair in Molecular Virology
01634 202957
Medway School of Pharmacy,
Anson Building,
Central Avenue,
Chatham Maritime,
Kent, ME4 4TB
United Kingdom
APA
Click to copy
Genova, C., Sampson, A., Scott, S., Cantoni, D., Neto, M., Bentley, E., … Temperton, N. J. (2021). Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes. Bio-Protocol.
Chicago/Turabian
Click to copy
Genova, C., A. Sampson, S. Scott, D. Cantoni, M. Neto, E. Bentley, G. Mattiuzzo, et al. “Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes.” Bio-protocol (2021).
MLA
Click to copy
Genova, C., et al. “Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes.” Bio-Protocol, 2021.
BibTeX Click to copy
@article{c2021a,
title = {Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes.},
year = {2021},
journal = {Bio-protocol},
author = {Genova, C. and Sampson, A. and Scott, S. and Cantoni, D. and Neto, M. and Bentley, E. and Mattiuzzo, G. and Wright, E. and Derveni, Mariliza and Auld, B. and Ferrara, Billy and Harrison, Dale and Said, Mohamed and Selim, Arwa Z. and Thompson, Elinor P. and Thompson, C. and Carnell, G. and Temperton, Nigel J.}
}
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.