An in silico and in vivo analysis of a novel multi-epitope peptide vaccine against hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is becoming more prevalent, making it one of the most serious worldwide health challenges. Several studies have focused on designing effective multi-epitope peptide vaccines against HCC in recent years. An in silico approach was used in prior investigations to create a multi-epitope peptide vaccine against HCC. Glypican-3 (GPC-3), melanoma-associated antigen-C2 (MAGE-C2), aspartyl-β-hydroxylase (ASPH), and New York esophageal squamous cell carcinoma 1 (NY-ESO-1) are four overexpressed antigens in HCC patients that we used in the vaccine's design. A multi-epitope peptide vaccine against HCC was designed through in silico antigen selection, physicochemical characterization, epitope prediction, structural analysis, cloning optimization, immune simulation, bacterial expression, protein purification, and immunological evaluation in rats. Four B-cells, nine MHC-I restricted, and eleven MHC-II restricted epitopes were selected based on their antigenicity score >0.5 and non-allergenicity with an adjuvant that included a segment of the microbial heat shock protein (HSP70) peptide 407-426 for vaccine construction. The relevant linkers were used to link each of the vaccine components and the cloned-in vector. This vector was successfully transformed into the Escherichia coli strain to further evaluate itsimmunological response and efficacy. The primary aim of this study is to design a vaccine and subsequently conduct laboratory experiments to evaluate its efficacy and safety profiles. It covers 90% of the global population, hence it is very effective. The vaccine's easily soluble nature was shown by its physicochemical properties, such as its hydropathicity index (-0.457) and aliphatic index (75.63). The vaccine has a high probability of being soluble in E. coli, with a solubility of 0.680. The ongoing results will provide insights into the efficacy of the vaccine against HCC.