The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression
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...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-12-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/11/12/1812 |
| _version_ | 1797506465453309952 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Mariano Maffei |
| collection | DOAJ |
| description | 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: <i>Escherichia coli</i>, 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. |
| first_indexed | 2024-03-10T04:33:02Z |
| format | Article |
| id | doaj.art-af5296f4183347728d8f456fd9719f2b |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T04:33:02Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-af5296f4183347728d8f456fd9719f2b2023-11-23T03:59:31ZengMDPI AGBiomolecules2218-273X2021-12-011112181210.3390/biom11121812The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous ExpressionMariano Maffei0Linda Celeste Montemiglio1Grazia Vitagliano2Luigi Fedele3Shaila Sellathurai4Federica Bucci5Mirco Compagnone6Valerio Chiarini7Cécile Exertier8Alessia Muzi9Giuseppe Roscilli10Beatrice Vallone11Emanuele Marra12Evvivax Biotech, Via di Castel Romano 100, 00128 Rome, ItalyInstitute 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, ItalyTakis Biotech, Via di Castel Romano 100, 00128 Rome, ItalyTakis Biotech, Via di Castel Romano 100, 00128 Rome, ItalyTakis Biotech, Via di Castel Romano 100, 00128 Rome, ItalyTakis Biotech, Via di Castel Romano 100, 00128 Rome, ItalyNeomatrix Biotech, Via di Castel Romano 100, 00128 Rome, ItalyTakis Biotech, Via di Castel Romano 100, 00128 Rome, ItalyDepartment of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza, University of Rome, P. le Aldo Moro, 5, 00185 Rome, ItalyTakis Biotech, Via di Castel Romano 100, 00128 Rome, ItalyEvvivax Biotech, Via di Castel Romano 100, 00128 Rome, ItalyDepartment of Biochemical Sciences “Alessandro Rossi Fanelli”, Sapienza, University of Rome, P. le Aldo Moro, 5, 00185 Rome, ItalyEvvivax Biotech, Via di Castel Romano 100, 00128 Rome, ItalyCOVID-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: <i>Escherichia coli</i>, 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.https://www.mdpi.com/2218-273X/11/12/1812SARS-CoV-2receptor-binding domainCOVID-19spike proteinheterologous expressionprotein production |
| spellingShingle | 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 The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression Biomolecules SARS-CoV-2 receptor-binding domain COVID-19 spike protein heterologous expression protein production |
| title | The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression |
| title_full | The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression |
| title_fullStr | The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression |
| title_full_unstemmed | The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression |
| title_short | The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression |
| title_sort | nuts and bolts of sars cov 2 spike receptor binding domain heterologous expression |
| topic | SARS-CoV-2 receptor-binding domain COVID-19 spike protein heterologous expression protein production |
| url | https://www.mdpi.com/2218-273X/11/12/1812 |
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