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Hybrid Proteins with Short Conformational Epitopes of the Receptor-Binding Domain of SARS-CoV-2 Spike Protein Promote Production of Virus-Neutralizing Antibodies When Used for Immunizationстатья
- Авторы: Karyagina Anna S., Gromov Alexander V., Grunina Tatyana M., Lyaschuk Alexander M., Poponova Maria S., Kleymenov Denis A., Strukova Natalia V., Generalova Maria S., Ryazanova Anna V., Galushkina Zoya M., Dobrynina Olga Yu, Bolshakova Tatyana N., Sergeeva Maria V., Romanovskaya-Romanko Ekaterina A., Krasilnikov Igor V., Subbotina Marina E., Lunin Vladimir G.
- Журнал: Biochemistry (Moscow)
- Том: 87
- Номер: 4
- Год издания: 2022
- Издательство: Pleiades Publishing, Ltd
- Местоположение издательства: Road Town, United Kingdom
- Первая страница: 319
- Последняя страница: 330
- DOI: 10.1134/s0006297922040022
- Аннотация: Based on the previously developed approach, hybrid recombinant proteins containing short conformational epitopes (a.a. 144 - 153, 337 - 346, 414 - 425, 496 - 507) of the receptorbinding domain (RBD) of SARSCoV2 Spike protein (S protein) were synthesized in Escherichia coli cells as potential components of epitope vaccines. Selected epitopes are involved in protein–protein interactions in the S protein complexes with neutralizing antibodies and ACE2 (angiotensin converting enzyme 2). The recombinant proteins were used for immunization of mice (three doses with 2 week intervals), and the immunogenicity of protein antigens and ability of the resulting sera to interact with inactivated SARSCoV2 and RBD produced in eukaryotic cells were examined. All recombinant proteins showed high immunogenicity; the highest titer in the RBD binding assay was demonstrated by the serum obtained after immunization with the protein containing epitope414 - 425. At the same time, the titers of sera obtained against other proteins in the RBD and inactivated virus binding assays were significantly lower than the titers of sera obtained with the previously produced four proteins containing the looplike epitopes 452 - 494 and 470 - 491, the conformation of which was fixed with a disulfide bond. We also studied activation of cell mediated immunity by the recombinant proteins that was monitored as changes in the levels of cytokines in the splenocytes of immunized mice. The most pronounced increase in the cytokine synthesis was observed in response to the proteins containing epitopes with disulfide bonds (452494, 470491), as well as epitopes 414 - 425 and 496 - 507. For some recombinant proteins with short conformational epitopes, adjuvant optimization allowed to obtained mouse sera displaying virusneutralizing activity in the microneutralization assay with live SARSCoV2 hCoV19/Russia/StPetersburg3524/2020 EPI_ISL_415710 GISAID). The results obtained can be used to develop epitope vaccines for prevention of COVID19 and other viral infections.
- Добавил в систему: Карягина-Жулина Анна Станиславовна