Abstract
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.
Highlights
At the end of 2019, a novel respiratory pathogen responsible for the COVID-19 disease, namely, severe acute respiratory syndrome-related coronavirus (SARS-CoV-2), emerged in Wuhan, China [1]
The Western blot analysis indicated that the protein was efficiently recognized by anti-His and anti-SARS-CoV-2 Spike S1 subunit antibodies (Figure 1c)
1.25 mg of purified receptor-binding domain (RBD) was obtained starting from 0.5 L of bacterial culture
Summary
At the end of 2019, a novel respiratory pathogen responsible for the COVID-19 disease, namely, severe acute respiratory syndrome-related coronavirus (SARS-CoV-2), emerged in Wuhan, China [1]. The virus spread worldwide causing one of largest outbreak of the century that rapidly progressed into a pandemic with more than 244 million of confirmed cases and 496 million deaths as of October 27th [2]. In response to this exceptional situation, an enormous effort has been made by the scientific community to study and characterize the pathogen and to quickly develop safe and effective prophylactic and therapeutic drugs. The 140 kDa SARS-CoV-2 S protein is organized into two major subunits
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