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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
- Source :
- Diagnostics, Vol 11, Iss 1808, p 1808 (2021), Diagnostics, Volume 11, Issue 10
- Publication Year :
- 2021
- Publisher :
- MDPI AG, 2021.
-
Abstract
- 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.
- Subjects :
- 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
Subjects
Details
- Language :
- English
- ISSN :
- 20754418
- Volume :
- 11
- Issue :
- 1808
- Database :
- OpenAIRE
- Journal :
- Diagnostics
- Accession number :
- edsair.doi.dedup.....2a32ddf0b49abba9d1cf6a8b74684dfe