This study aimed to design a multi-epitope peptide vaccine targeting prostate-specific membrane antigen (PSMA), six-transmembrane epithelial antigen of prostate 1 (STEAP1), and B7-H3, three biomarkers strongly associated with prostate cancer progression.<h4>Methods</h4>A multi-layered immunoinformatics-driven approach was employed, integrating epitope prediction, antigenicity and immunogenicity assessment, allergenicity and toxicity screening, population coverage analysis, molecular docking, and molecular dynamics simulations. The gene discussed is FOLH1; the disease is prostate cancer.