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

doi:10.3390/biom11121812

Biomolecules (Dec 2021)

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

DOI: https://doi.org/10.3390/biom11121812
Journal volume & issue: Vol. 11, no. 12
p. 1812

Abstract

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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.

Published in Biomolecules

ISSN: 2218-273X (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: https://www.mdpi.com/journal/biomolecules

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