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Publications

Journal Article Epublish

PubMed: 40558387 DOI: 10.3390/biotech14020038

Strategy for the Construction of SARS-CoV-2 S and N Recombinant Proteins and Their Immunogenicity Evaluation.

BioTech (Basel) 14(2)

Authors: 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

Affiliations

  • Post-Graduate Program in Microbiology, Department of Microbiology, State University of Londrina, Londrina 86.055-900, Brazil.
  • Laboratory of Molecular Biology of Microorganisms, Department of Microbiology, State University of Londrina, Londrina 86.055-900, Brazil.
  • Carlos Chagas Institute, Oswaldo Cruz Foundation (FIOCRUZ-PR), Curitiba 81350-010, Brazil.
  • Post-Graduate Program in Clinical and Laboratory Physiopathology, State University of Londrina, Londrina 86.055-900, Brazil.
  • Department of Immunology, Parasitology and General Pathology, State University of Londrina, Londrina 86.055-900, Brazil.
  • Institute of Molecular Biology of Paraná, Curitiba 81350-010, Brazil.
  • Laboratory of Technological Innovation in the Development of Pharmaceuticals and Cosmetics, State University of Maringá, Maringá 87020-900, Brazil.
  • Department of Medicine, Pontifical Catholic University of Paraná, Londrina 86067-000, Brazil.

Abstract

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 Escherichia coli 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.

MeSH

Sars-Cov-2, Epitopes, Covid-19, Synthetic Biology, Severe Acute Respiratory Syndrome-Related Coronavirus, Recombinant Proteins