The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression
Mariano Maffei,
Linda Celeste Montemiglio,
Grazia Vitagliano,
Luigi Fedele,
Shaila Sellathurai,
Federica Bucci,
Mirco Compagnone,
Valerio Chiarini,
Cécile Exertier,
Alessia Muzi,
Giuseppe Roscilli,
Beatrice Vallone,
Emanuele Marra
Affiliations
Mariano Maffei
Evvivax Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Linda Celeste Montemiglio
Institute of Molecular Biology and Pathology (IBPM), National Research Council, c/o Department of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza, University of Rome, P. le Aldo Moro, 5, 00185 Rome, Italy
Grazia Vitagliano
Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Luigi Fedele
Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Shaila Sellathurai
Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Federica Bucci
Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Mirco Compagnone
Neomatrix Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Valerio Chiarini
Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Cécile Exertier
Department of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza, University of Rome, P. le Aldo Moro, 5, 00185 Rome, Italy
Alessia Muzi
Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Giuseppe Roscilli
Evvivax Biotech, Via di Castel Romano 100, 00128 Rome, Italy
Beatrice Vallone
Department of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza, University of Rome, P. le Aldo Moro, 5, 00185 Rome, Italy
Emanuele Marra
Evvivax Biotech, Via di Castel Romano 100, 00128 Rome, Italy
COVID-19 is a highly infectious disease caused by a newly emerged coronavirus (SARS-CoV-2) that has rapidly progressed into a pandemic. This unprecedent emergency has stressed the significance of developing effective therapeutics to fight the current and future outbreaks. The receptor-binding domain (RBD) of the SARS-CoV-2 surface Spike protein is the main target for vaccines and represents a helpful “tool” to produce neutralizing antibodies or diagnostic kits. In this work, we provide a detailed characterization of the native RBD produced in three major model systems: Escherichia coli, insect and HEK-293 cells. Circular dichroism, gel filtration chromatography and thermal denaturation experiments indicated that recombinant SARS-CoV-2 RBD proteins are stable and correctly folded. In addition, their functionality and receptor-binding ability were further evaluated through ELISA, flow cytometry assays and bio-layer interferometry.
