Borges Paulo Henrique Guilherme, Gregio Barbara, Suzukawa Helena Tiemi, Silva-Rodrigues Gislaine, Andreassa Emanuella de Castro, Castro Isabela Madeira de, Bartolomeu-Gonçalves Guilherme, Venancio Emerson José, Pinge-Filho Phileno, Góes Viviane Monteiro, Nakamura Celso Vataru, Tavares Eliandro Reis, Souza Tatiana de Arruda Campos Brasil de, Yamada-Ogatta Sueli Fumie, Yamauchi Lucy Megumi
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.
BioTech (Basel). 2025 May 23;14(2):38. doi: 10.3390/biotech14020038.
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 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.
本研究报告了含有免疫显性表位的合成严重急性呼吸综合征冠状病毒2（SARS-CoV-2）刺突（S）蛋白和核蛋白（N）的构建、表达及纯化。pET28aS_epit构建体包含表位287 - 317、402、507、524 - 598和601 - 640，而pET28aN_epit构建体包含残基42 - 62、153 - 172和355 - 401。两种蛋白的商业序列用作对照。使用BL21(DE3) star菌株在37°C下表达这四种构建体。结果表明，与N蛋白构建体不同，S蛋白构建体不溶。两种重组蛋白均能在小鼠中诱导免疫反应，并被来自COVID-19阳性和/或接种SARS-CoV-2疫苗的人类血清中的抗体识别。在我们的构建体和市售蛋白之间未观察到免疫识别的显著差异。总之，S_epit和N_epit可能是基于免疫反应开发控制SARS-CoV-2感染新策略的有前景的起始点。
