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.
The nuts and bolts of SARS-CoV-2 spike receptor-binding domain heterologous expression / 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. - In: BIOMOLECULES. - ISSN 2218-273X. - 11:12(2021), p. 1812. [10.3390/biom11121812]
The nuts and bolts of SARS-CoV-2 spike receptor-binding domain heterologous expression
Montemiglio, Linda Celeste;Roscilli, Giuseppe;Vallone, Beatrice;
2021
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.| File | Dimensione | Formato | |
|---|---|---|---|
|
Maffei_The Nuts_2021.pdf.pdf
accesso aperto
Tipologia:
Versione editoriale (versione pubblicata con il layout dell'editore)
Licenza:
Creative commons
Dimensione
2.46 MB
Formato
Adobe PDF
|
2.46 MB | Adobe PDF |
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
