Background and Aim: Cervical cancer, primarily caused by human papillomavirus (HPV), remains a significant health challenge. High-risk HPV types, particularly through the persistent expression of oncoproteins E6 and E7, disrupt tumor suppressor functions. Recognizing the role of Heat Shock Protein 110 (HSP110) in enhancing immune responses, this study explores the design and therapeutic potential of a novel T-cell-epitope-based DNA vaccine targeting these viral components. Materials and Methods: We constructed a chimeric HPV16 E6/E7/HSP110 vaccine with high molecular weight, a basic nature, good aqueous solubility, and prolonged stability/half-life. We investigated immune reactivity, conservancy among HPV strains, and affinity to certain HLA types for potential antigenic peptides emanating from E6, E7, and HSP110. To this end, the three-dimensional structure of this protein was further refined using server-based methods and its interaction with HLA class I [HLA-A11:01] and MHC-II molecules [HLA-DRB1_01:01] was assessed by in silico molecular docking studies. Additionally, we performed a cell-based ELISA on HPV16-transformed TC-1 target cells using sera from healthy vaccinated mice to assess antigen-specific binding to cellular targets; cytokines were quantified separately by sandwich ELISA on culture supernatants as a complementary readout. Results: The T-cell epitopes of HPV16 E6, E7, and HSP110 were predicted, and 10 epitopes with strong antigenic properties and non-toxicity were selected. These epitopes were linked using a KK linker to generate a multi-epitope vaccine. The resulting vaccine exhibited high antigenicity, non-allergenicity, and good solubility with favorable physicochemical properties. The 3D structure of the vaccine and its interactions with HLA class I [HLA-A11:01] and class II [HLA-DRB1_01:01] were successfully demonstrated. Molecular dynamics (MD) simulations confirmed the stability of the vaccine binding to HLA-A11:01. Codon optimization and virtual cloning predicted high expression in E. coli K12; in vitro, Western blotting in HEK293T cells confirmed expression of the chimeric construct. Preliminary cell-ELISA indicated increased secretion of selected cytokines (e.g., IFN-?, IL-2, TNF-?) relative to appropriate controls, supporting cell based immunogenicity. Conclusion: The data collected demonstrated that the proposed construct represents a novel and promising therapeutic vaccine candidate for cervical cancer. Although further validation with additional experimental work and clinical trials is needed, our study provides a starting point to guide the investigation of new treatment strategies for cervical cancer.