@article{Di_Genova_C-2021_151100,
title = {Production, titration, neutralisation, storage and lyophilisation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lentiviral pseudotypes},
author = {Di 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, M. and Auld, B. and Ferrara, B. and Harrison, D. and Said, M. and Selim, A. and Thompson, E. and Thompson, C. and Carnell, G. and Temperton, N.},
month = {nov},
year = {2021},
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.},
volume = {11},
issue = {21},
journal = {Bio-Protocol Journal},
publisher = {Bio-Protocol},
url = {https://doi.org/10.21769/BioProtoc.4236},
}
