Our first paper on the study of SARS-CoV-2 is finally out. This study developed at BioSIM (UCIBIO@REQUIMTE – Faculdade de Medicina, Universidade do Porto) and at University of Bordeaux and University of Nantes was published in Viruses (IF 3.816) and resulted from a collaboration initiated through the Crowdfight COVID19 initiative (https://crowdfightcovid19.org).
In silico, in vitro and in cellulo models for monitoring SARS-CoV-2 spike/human ACE2 complex, viral entry and cell fusion
DOI: 10.3390/v13030365 | Viruses
Our study presents a combination of in silico, in vitro and in cellulo models for monitoring SARS-CoV-2 spike/human ACE2 complex, viral entry and cell fusion. The study reports a novel computational model of the SARS-CoV-2 S/ACE2 complex as well as its biochemical and biophysical monitoring using pulldown, AlphaLISA and biolayer interferometry (BLI) binding assays. This led us to determine the kinetic parameters of the S/ACE2 association and dissociation steps. In parallel to these in vitro approaches, we developed in cellulo transduction assays using SARS-CoV-2 pseudotyped lentiviral vectors and HEK293T-ACE2 cell lines generated in-house. This allowed us to recapitulate the early replication stage of the infection mediated by the S/ACE2 interaction and to detect cell fusion induced by the interaction. Finally, a cell imaging system was set up to directly monitor the S/ACE2 interaction in a cellular context, and a flow cytometry assay was developed to quantify this association at the cell surface.
These combined models have been continuously applied through the last year at the Universities of Porto, Bordeaux and Nantes in the evaluation and characterization of the anti-viral activity against SARS-CoV-2 of thousands of molecules, enabling the identification of several promising molecules for drug development.