Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
| Autor: | Carlos E. Miguel-Rodríguez, Jose L. Maravillas Montero, José J. Torres-Ruíz, Leopoldo Santos Argumedo, Juan M. Alvarado-Orozco, Daniela Roa-Velázquez, Leticia Cedillo-Barrón, Erik Saul Sanchez-Salguero, David Andrés Fernández-Benavides, Sandra Paola Martínez-Frías, Edgar Morales-Ríos, Jessica G. Filisola-Villaseñor, Julio García-Cordero, Juvenal Mendoza-Ramírez, Diana Gómez-Martín |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Medicine (General)
receptor binding domain Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Protein subunit viruses Clinical Biochemistry nucleocapsid Spike Protein Biology spike protein Virology Article law.invention Serology Plasmid R5-920 Antigen coronavirus disease law Recombinant DNA biology.protein Antibody severe acute respiratory syndrome coronavirus 2 |
| Zdroj: | Diagnostics, Vol 11, Iss 1808, p 1808 (2021) Diagnostics Volume 11 Issue 10 |
| ISSN: | 2075-4418 |
| Popis: | The coronavirus disease 2019 (COVID-19) pandemic has reached an unprecedented level. There is a strong demand for diagnostic and serological supplies worldwide, making it necessary for countries to establish their own technologies to produce high-quality biomolecules. The two main viral antigens used for the diagnostics for severe acute respiratory syndrome coronavirus (SARS-CoV-2) are the structural proteins spike (S) protein and nucleocapsid (N) protein. The spike protein of SARS-CoV-2 is cleaved into S1 and S2, in which the S1 subunit has the receptor-binding domain (RBD), which induces the production of neutralizing antibodies, whereas nucleocapsid is an ideal target for viral antigen-based detection. In this study, we designed plasmids, pcDNA3.1/S1 and pcDNA3.1/N, and optimized their expression of the recombinant S1 and N proteins from SARS-CoV-2 in a mammalian system. The RBD was used as a control. The antigens were successfully purified from Expi293 cells, with high yields of the S1, N, and RBD proteins. The immunogenic abilities of these proteins were demonstrated in a mouse model. Further, enzyme-linked immunosorbent assays with human serum samples showed that the SARS-CoV-2 antigens are a suitable alternative for serological assays to identify patients infected with COVID-19. |
| Databáze: | OpenAIRE |
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