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Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications

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 use...

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Autores principales: García-Cordero, Julio, Mendoza-Ramírez, Juvenal, Fernández-Benavides, David, Roa-Velazquez, Daniela, Filisola-Villaseñor, Jessica, Martínez-Frías, Sandra Paola, Sanchez-Salguero, Erik Saul, Miguel-Rodríguez, Carlos E., Maravillas Montero, Jose L., Torres-Ruiz, Jose J., Gómez-Martín, Diana, Argumedo, Leopoldo Santos, Morales-Ríos, Edgar, Alvarado-Orozco, Juan M., Cedillo-Barrón, Leticia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8534734/
https://www.ncbi.nlm.nih.gov/pubmed/34679506
http://dx.doi.org/10.3390/diagnostics11101808
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author García-Cordero, Julio
Mendoza-Ramírez, Juvenal
Fernández-Benavides, David
Roa-Velazquez, Daniela
Filisola-Villaseñor, Jessica
Martínez-Frías, Sandra Paola
Sanchez-Salguero, Erik Saul
Miguel-Rodríguez, Carlos E.
Maravillas Montero, Jose L.
Torres-Ruiz, Jose J.
Gómez-Martín, Diana
Argumedo, Leopoldo Santos
Morales-Ríos, Edgar
Alvarado-Orozco, Juan M.
Cedillo-Barrón, Leticia
author_facet García-Cordero, Julio
Mendoza-Ramírez, Juvenal
Fernández-Benavides, David
Roa-Velazquez, Daniela
Filisola-Villaseñor, Jessica
Martínez-Frías, Sandra Paola
Sanchez-Salguero, Erik Saul
Miguel-Rodríguez, Carlos E.
Maravillas Montero, Jose L.
Torres-Ruiz, Jose J.
Gómez-Martín, Diana
Argumedo, Leopoldo Santos
Morales-Ríos, Edgar
Alvarado-Orozco, Juan M.
Cedillo-Barrón, Leticia
author_sort García-Cordero, Julio
collection PubMed
description 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.
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spelling pubmed-85347342021-10-23 Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications García-Cordero, Julio Mendoza-Ramírez, Juvenal Fernández-Benavides, David Roa-Velazquez, Daniela Filisola-Villaseñor, Jessica Martínez-Frías, Sandra Paola Sanchez-Salguero, Erik Saul Miguel-Rodríguez, Carlos E. Maravillas Montero, Jose L. Torres-Ruiz, Jose J. Gómez-Martín, Diana Argumedo, Leopoldo Santos Morales-Ríos, Edgar Alvarado-Orozco, Juan M. Cedillo-Barrón, Leticia Diagnostics (Basel) Article 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. MDPI 2021-09-30 /pmc/articles/PMC8534734/ /pubmed/34679506 http://dx.doi.org/10.3390/diagnostics11101808 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
García-Cordero, Julio
Mendoza-Ramírez, Juvenal
Fernández-Benavides, David
Roa-Velazquez, Daniela
Filisola-Villaseñor, Jessica
Martínez-Frías, Sandra Paola
Sanchez-Salguero, Erik Saul
Miguel-Rodríguez, Carlos E.
Maravillas Montero, Jose L.
Torres-Ruiz, Jose J.
Gómez-Martín, Diana
Argumedo, Leopoldo Santos
Morales-Ríos, Edgar
Alvarado-Orozco, Juan M.
Cedillo-Barrón, Leticia
Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
title Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
title_full Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
title_fullStr Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
title_full_unstemmed Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
title_short Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications
title_sort recombinant protein expression and purification of n, s1, and rbd of sars-cov-2 from mammalian cells and their potential applications
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8534734/
https://www.ncbi.nlm.nih.gov/pubmed/34679506
http://dx.doi.org/10.3390/diagnostics11101808
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