Biomolecules · 2021
The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Maffei Mariano, Montemiglio Linda Celeste, Vitagliano Grazia, Fedele Luigi, Sellathurai Shaila, Bucci Federica, Compagnone Mirco, Chiarini Valerio, Exertier Cécile, Muzi Alessia, Roscilli Giuseppe, Vallone Beatrice, Marra Emanuele
Аннотация
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.