Production, titration, neutralisation and storage of SARS-CoV-2 lentiviral pseudotypes

Di Genova, Cecilia; Sampson, Alex; Scott, Simon; Cantoni, Diego; Mayora-Neto, Martin; Bentley, Emma; Mattiuzzo, Giada; Wright, Edward; Derveni, Mariliza; Auld, Bethany; Ferrara, Bill; Harrison, Dale; Said, Mohamed; Selim, Arwa; Thompson, Elinor; Thompson, Craig; Carnell, George; Temperton, Nigel

doi:10.6084/m9.figshare.13502580.v2

Temperton SARS-CoV-2 FS.pdf (2.36 MB)

Production, titration, neutralisation and storage of SARS-CoV-2 lentiviral pseudotypes

Version 2 2020-12-30, 12:54

Version 1 2020-12-30, 12:01

preprint

posted on 2020-12-30, 12:54 authored by Cecilia Di Genova, Alex Sampson, Simon Scott, Diego Cantoni, Martin Mayora-Neto, Emma Bentley, Giada Mattiuzzo, Edward WrightEdward Wright, Mariliza Derveni, Bethany Auld, Bill Ferrara, Dale Harrison, Mohamed Said, Arwa Selim, Elinor Thompson, Craig Thompson, George Carnell, Nigel TempertonNigel Temperton

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.