Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation
This study reports the construction, expression, and purification of synthetic SARS-CoV-2 spike (S) and nucleoprotein (N) containing immunodominant epitopes. The pET28aS_epit construct included epitopes 287-317, 402, 507, 524-598, and 601-640, while the pET28aN_epit construct included residues 42-62, 153-172, and 355-401. Commercial sequences of both proteins were used as controls. The four constructs were expressed using the <i>Escherichia coli</i> BL21(DE3) star strain at 37 °C. The results show that the S protein constructs were insoluble, unlike the N protein constructs. Both recombinant proteins induced immune responses in mice and were recognized by antibodies present in sera from COVID-19-positive and/or SARS-CoV-2-vaccinated humans. No significant differences in immune recognition were observed between our constructs and the commercially available proteins. In conclusion, S_epit and N_epit could be promising starting points for the development of new strategies based on immunological reactions for the control of SARS-CoV-2 infections.
Authors
- Guilherme Bartolomeu Gonçalves (ORCID: https://orcid.org/0000-0002-3497-5362)
- Celso Vataru Nakamura (ORCID: https://orcid.org/0000-0002-9911-7369)
- Isabela Madeira de Castro (ORCID: https://orcid.org/0009-0008-2933-5783)
- Emanuella de Castro Andreassa (ORCID: https://orcid.org/0000-0002-3297-5529)
- Eliandro Reis Tavares (ORCID: https://orcid.org/0000-0001-6234-1589)
- Helena Tiemi Suzukawa (ORCID: https://orcid.org/0000-0002-2694-6553)
- Emerson José Venâncio (ORCID: https://orcid.org/0000-0002-3735-9207)
- Viviane Monteiro Góes
- Tatiana de Arruda Campos Brasil de Souza (ORCID: https://orcid.org/0000-0002-1850-6080)
- Sueli Fumie Yamada‐Ogatta (ORCID: https://orcid.org/0000-0003-1183-146X)
- Lucy Megumi Yamauchi (ORCID: https://orcid.org/0000-0003-2241-8814)
- Phileno Pinge‐Filho (ORCID: https://orcid.org/0000-0002-1500-3998)
- Paulo Henrique Guilherme Borges (ORCID: https://orcid.org/0000-0002-5412-053X)
- Gislaine Silva-Rodrigues
- Barbara Gregio
Institutions
- Universidade Estadual de Maringá (BR)
- Universidade Estadual de Londrina (BR)
- Pontifícia Universidade Católica do Paraná (BR)
- Instituto de Biologia Molecular do Paraná (BR)
- Fundação Oswaldo Cruz (BR)
Publication Details
- Journal
- BioTech
- Published
- 2025-05-23
- DOI
- https://doi.org/10.3390/biotech14020038
- Primary Topic
- SARS-CoV-2 and COVID-19 Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
- Conselho Nacional de Desenvolvimento Científico e Tecnológico
- Fundação Araucária
- Financiadora de Estudos e Projetos