Immunoinformatics-Based Design of a Multiepitope Vaccine Candidate Against Jembrana Disease Virus: Protein Expression in Escherichia coli BL۲۱, Molecular Docking, and Molecular Dynamics Simulation

Jembrana disease virus (JDV) remains a major threat to Bali cattle, while currently available inactivated vaccines provide only limited and short-term protection. This study applied an integrated immunoinformatics and molecular simulation approach to design and evaluate a multiepitope vaccine candidate against JDV. B-cell, cytotoxic T lymphocyte (CTL), and helper T lymphocyte (HTL) epitopes were predicted from the capsid (CA) and transmembrane (TM) proteins and assembled into a ۳۰۹-amino-acid construct incorporating the ۵۰S ribosomal protein L۷/L۱۲ as a TLR۴-targeting adjuvant. Structural modeling and validation confirmed the reliability and stereochemical quality of the three-dimensional structure. Physicochemical and immunological analyses indicated that the construct was stable, hydrophilic, antigenic, non-allergenic, and non-toxic. Molecular docking demonstrated favorable binding affinity between the vaccine construct and the TLR۴ receptor. Molecular dynamics simulations further supported complex stability, as reflected by acceptable RMSF values and limited residue fluctuation at the binding interface. In silico immune simulations predicted strong humoral and cellular immune responses, characterized by elevated IgM and IgG antibody titers, expansion of B- and T-lymphocyte populations, and the formation of memory cells following booster administrations. Codon optimization and in silico cloning confirmed compatibility with the pET-۲۸b (+) expression system. Recombinant protein expression in Escherichia coli BL۲۱ (DE۳) was validated by molecular analysis and SDS–PAGE, revealing a protein of approximately ۳۵.۱۷ kDa predominantly in soluble form.These findings highlight the potential of the proposed multiepitope construct as a promising JDV vaccine candidate and support further experimental validation.

کلیدواژه ها:

Immunoinformatics ، Jembrana disease virus ، Molecular docking ، Molecular Dynamics Simulation ، multiepitope vaccine ، recombinant protein expression