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...
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MDPI AG
2025-05-01
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| Online Access: | https://www.mdpi.com/2673-6284/14/2/38 |
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| author | Paulo Henrique Guilherme Borges Barbara Gregio Helena Tiemi Suzukawa Gislaine Silva-Rodrigues Emanuella de Castro Andreassa Isabela Madeira de Castro Guilherme Bartolomeu-Gonçalves Emerson José Venancio Phileno Pinge-Filho Viviane Monteiro Góes Celso Vataru Nakamura Eliandro Reis Tavares Tatiana de Arruda Campos Brasil de Souza Sueli Fumie Yamada-Ogatta Lucy Megumi Yamauchi |
| author_facet | Paulo Henrique Guilherme Borges Barbara Gregio Helena Tiemi Suzukawa Gislaine Silva-Rodrigues Emanuella de Castro Andreassa Isabela Madeira de Castro Guilherme Bartolomeu-Gonçalves Emerson José Venancio Phileno Pinge-Filho Viviane Monteiro Góes Celso Vataru Nakamura Eliandro Reis Tavares Tatiana de Arruda Campos Brasil de Souza Sueli Fumie Yamada-Ogatta Lucy Megumi Yamauchi |
| author_sort | Paulo Henrique Guilherme Borges |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-c0c01844a2f847c9a8b034c8c8e519db |
| institution | OA Journals |
| issn | 2673-6284 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | BioTech |
| spelling | doaj-art-c0c01844a2f847c9a8b034c8c8e519db2025-08-20T02:24:26ZengMDPI AGBioTech2673-62842025-05-011423810.3390/biotech14020038Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity EvaluationPaulo Henrique Guilherme Borges0Barbara Gregio1Helena Tiemi Suzukawa2Gislaine Silva-Rodrigues3Emanuella de Castro Andreassa4Isabela Madeira de Castro5Guilherme Bartolomeu-Gonçalves6Emerson José Venancio7Phileno Pinge-Filho8Viviane Monteiro Góes9Celso Vataru Nakamura10Eliandro Reis Tavares11Tatiana de Arruda Campos Brasil de Souza12Sueli Fumie Yamada-Ogatta13Lucy Megumi Yamauchi14Post-Graduate Program in Microbiology, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilPost-Graduate Program in Microbiology, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilLaboratory of Molecular Biology of Microorganisms, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilPost-Graduate Program in Microbiology, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilCarlos Chagas Institute, Oswaldo Cruz Foundation (FIOCRUZ-PR), Curitiba 81350-010, BrazilPost-Graduate Program in Microbiology, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilPost-Graduate Program in Clinical and Laboratory Physiopathology, State University of Londrina, Londrina 86.055-900, BrazilDepartment of Immunology, Parasitology and General Pathology, State University of Londrina, Londrina 86.055-900, BrazilDepartment of Immunology, Parasitology and General Pathology, State University of Londrina, Londrina 86.055-900, BrazilInstitute of Molecular Biology of Paraná, Curitiba 81350-010, BrazilLaboratory of Technological Innovation in the Development of Pharmaceuticals and Cosmetics, State University of Maringá, Maringá 87020-900, BrazilLaboratory of Molecular Biology of Microorganisms, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilCarlos Chagas Institute, Oswaldo Cruz Foundation (FIOCRUZ-PR), Curitiba 81350-010, BrazilLaboratory of Molecular Biology of Microorganisms, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilLaboratory of Molecular Biology of Microorganisms, Department of Microbiology, State University of Londrina, Londrina 86.055-900, BrazilThis 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.https://www.mdpi.com/2673-6284/14/2/38SARS-CoV-2immunoinformaticspurification of recombinant proteinssynthetic biology |
| spellingShingle | Paulo Henrique Guilherme Borges Barbara Gregio Helena Tiemi Suzukawa Gislaine Silva-Rodrigues Emanuella de Castro Andreassa Isabela Madeira de Castro Guilherme Bartolomeu-Gonçalves Emerson José Venancio Phileno Pinge-Filho Viviane Monteiro Góes Celso Vataru Nakamura Eliandro Reis Tavares Tatiana de Arruda Campos Brasil de Souza Sueli Fumie Yamada-Ogatta Lucy Megumi Yamauchi Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation BioTech SARS-CoV-2 immunoinformatics purification of recombinant proteins synthetic biology |
| title | Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation |
| title_full | Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation |
| title_fullStr | Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation |
| title_full_unstemmed | Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation |
| title_short | Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation |
| title_sort | strategy for the construction of sars cov 2 s and n recombinant proteins and their immunogenicity evaluation |
| topic | SARS-CoV-2 immunoinformatics purification of recombinant proteins synthetic biology |
| url | https://www.mdpi.com/2673-6284/14/2/38 |
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