Production, titration, neutralisation, storage and lyophilisation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lentiviral pseudotypes

Genova, C. et al. (2021) Production, titration, neutralisation, storage and lyophilisation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lentiviral pseudotypes. Bio-protocol, 11(21), e4236. (doi: 10.21769/bioprotoc.4236) (PMID:34859134) (PMCID:PMC8595416)

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: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Cantoni, Dr Diego
• Authors: Genova, C., Sampson, A., Scott, S., Cantoni, D., Neto, M., Bentley, E., Mattiuzzo, G., Wright, E., Derveni, M., Auld, B., Ferrara, B., Harrison, D., Said, M., Selim, A., Thompson, E., Thompson, C., Carnell, G., and Temperton, N.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Bio-protocol
• Publisher: Bio-Protocol
• ISSN: 2331-8325
• ISSN (Online): 2331-8325