Lentiviral Pseudotypes for SARS-CoV-2 Variant Assay
Lentiviral pseudotypes were produced by transient transfection of HEK293T/17 cells with packaging plasmids p8.91 (45 (link), 46 ) and pCSFLW (47 (link)) and different SARS-CoV-2 VOC spike-bearing expression plasmids using the Fugene-HD transfection reagent (48 (link), 49 (link)). Supernatants were harvested after 48h, passed through a 0.45 µm cellulose acetate filter and titrated on HEK293T/17 cells transiently expressing human ACE-2 and TMPRSS2. Target HEK293T/17 cells were transfected 24h prior with 2 µg pCAGGS-huACE-2 and 75 ng pCAGGS-TMPRSS2 (16 (link), 50 (link)).
Carnell G.W., Billmeier M., Vishwanath S., Suau Sans M., Wein H., George C.L., Neckermann P., Del Rosario J.M., Sampson A.T., Einhauser S., Aguinam E.T., Ferrari M., Tonks P., Nadesalingam A., Schütz A., Huang C.Q., Wells D.A., Paloniemi M., Jordan I., Cantoni D., Peterhoff D., Asbach B., Sandig V., Temperton N., Kinsley R., Wagner R, & Heeney J.L. (2023). Glycan masking of a non-neutralising epitope enhances neutralising antibodies targeting the RBD of SARS-CoV-2 and its variants. Frontiers in Immunology, 14, 1118523.
Different SARS-CoV-2 VOC spike-bearing expression plasmids
dependent variables
Lentiviral pseudotype titers
control variables
Packaging plasmids p8.91 and pCSFLW
Transfection of HEK293T/17 cells
Transfection of target HEK293T/17 cells with pCAGGS-huACE-2 and pCAGGS-TMPRSS2
positive controls
HEK293T/17 cells transiently expressing human ACE-2 and TMPRSS2
negative controls
Not explicitly mentioned
Annotations
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