The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression

Biomolecules

Beatrice Vallone¹  Cécile Exertier¹  Giuseppe Roscilli²  Mariano Maffei²  Emanuele Marra²  Linda Celeste Montemiglio³  Mirco Compagnone⁴  Shaila Sellathurai⁵  Luigi Fedele⁵  Federica Bucci⁵  Grazia Vitagliano⁵  Alessia Muzi⁵  Valerio Chiarini⁵ 

[1] Department of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza, University of Rome, P. le Aldo Moro, 5, 00185 Rome, Italy;Evvivax Biotech, Via di Castel Romano 100, 00128 Rome, Italy;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;Neomatrix Biotech, Via di Castel Romano 100, 00128 Rome, Italy;Takis Biotech, Via di Castel Romano 100, 00128 Rome, Italy;

关键词: SARS-CoV-2;    receptor-binding domain;    COVID-19;    spike protein;    heterologous expression;    protein production;   

DOI  :  10.3390/biom11121812

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown