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. 2024 Jul 18;14:16605. doi: 10.1038/s41598-024-67781-5

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Multiepitope polypeptide characteristics and in silico and in vitro safety validation. (A) Structural model of the multiepitope polypeptide obtained with I-TASSER. (B) In silico safety validation data for the polypeptide. (C) Evaluation of the viability of Vero-Dog-SLAM, MDCK and cPBMC cells after polypeptide treatment through the MTT assay. Cells were treated with twofold serial dilutions of polypeptide from 6.125 to 25 nM for 48 h. Cells without polypeptide were used as a viability negative control, and cells treated with 0.5% Triton X-100® served as a cytotoxicity positive control. (D) Hemolytic potential of the polypeptide in canine RBCs. PBS was used as a negative hemolytic control, and 0.1% Triton X-100® was used as a positive hemolytic control. Two independent experiments with 4 replicates (n = 8) were carried out, and nontreated cells were used as negative controls. The means and coefficients of variation are shown in the graph. *Evaluated with the Canis lupus familiaris proteome as the dataset. **Score of 0.57349, classified as a probable antigen MCH-I. ***Docking score obtained with the HPEPDOCK tool.