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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

Diagnostics, ISSN: 2075-4418, Vol: 11, Issue: 10
2021
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Article 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.

Bibliographic Details

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

MDPI AG

Biochemistry, Genetics and Molecular Biology

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