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

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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Main 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
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:BioTech
Subjects:
SARS-CoV-2
immunoinformatics
purification of recombinant proteins
synthetic biology
Online Access:https://www.mdpi.com/2673-6284/14/2/38
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Summary: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.
ISSN:2673-6284

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