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The present study aimed to construct a synthetic antigen multi-epitope SARS-CoV-2 to produce a DNA vaccine. Methods: A database of previously predicted Spike and Nucleocapsid protein epitopes was created, and these epitopes were analyzed for immunogenicity, conservation, population coverage, and molecular docking. Results: A synthetic antigen with 15 epitopes considered immunogenic, conserved even in the face of variants and that were able to anchor themselves in the appropriate HLA site, together had more than 90% worldwide coverage. A multi-epitope construct was developed with the sequences of these peptides separated from each other by linkers, cloned into the pVAX1 vector. This construct was evaluated in vivo as a DNA vaccine and elicited T CD4+ and T CD8+ cell expansion in the blood and spleen. In hematological analyses, there was an increase in lymphocytes, monocytes, and neutrophils between the two doses. Furthermore, based on histopathological analysis, the vaccines did not cause any damage to the organs analyzed. Conclusions: The present study generated a multi-epitope synthetic vaccine antigen capable of generating antibody-mediated and cellular immune responses.<\/jats:p>","DOI":"10.3390\/vaccines13020149","type":"journal-article","created":{"date-parts":[[2025,1,31]],"date-time":"2025-01-31T06:42:36Z","timestamp":1738305756000},"page":"149","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9360-1275","authenticated-orcid":false,"given":"Maria da Concei\u00e7\u00e3o Viana","family":"Inven\u00e7\u00e3o","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Studies and Experimental Therapy\u2014LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1660-4901","authenticated-orcid":false,"given":"Larissa Silva de","family":"Mac\u00eado","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Studies and Experimental Therapy\u2014LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0715-8674","authenticated-orcid":false,"given":"Ingrid Andr\u00eassa de","family":"Moura","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Studies and Experimental Therapy\u2014LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2089-6436","authenticated-orcid":false,"given":"Lucas Alexandre Barbosa de Oliveira","family":"Santos","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Genetics and Biotechnology (GMBio), Department of Biology, Center for Biological and Health Sciences, Federal University of Sergipe, S\u00e3o Crist\u00f3v\u00e3o 49100-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8185-9818","authenticated-orcid":false,"given":"Benigno Cristofer Flores","family":"Espinoza","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Studies and Experimental Therapy\u2014LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5576-3652","authenticated-orcid":false,"given":"Samara Sousa de","family":"Pinho","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Studies and Experimental Therapy\u2014LEMTE, Department of Genetics, Federal 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