TY - JOUR
AU - Di Genova, Cecilia
AU - Sampson, Alexander Thomas
AU - Scott, Simon
AU - Cantoni, Diego
AU - Neto, Martin
AU - Bentley, Emma M.
AU - Mattiuzzo, Giada
AU - Wright, Edward
AU - Derveni, Mariliza
AU - Auld, Bethany
AU - Ferrara, Bill
AU - Harrison, Dale
AU - Said, Mohamed
AU - Selim, Arwa
AU - Thompson, Elinor
AU - Thompson, Craig P.
AU - Carnell, George W.
AU - Temperton, Nigel
PY - 2021
DA - November
TI - Production, titration, neutralisation, storage and lyophilisation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lentiviral pseudotypes
JO - Bio-Protocol Journal
VL - 11
IS - 21
DO - https://doi.org/10.21769/BioProtoc.4236
AB - 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.
PB - Bio-Protocol
UR - https://eprints.gla.ac.uk/308334/
KW - Coronavirus (COVID-19)
ER