Immunoinformatics-Based Design of a Multiepitope Vaccine Candidate Against Jembrana Disease Virus: Protein Expression in Escherichia coli BL21, Molecular Docking, and Molecular Dynamics Simulation
نویسندگان
1 Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Gadjah Mada, Yogyakarta, 55281, Indonesia
2 Study Program of Doctor in Biotechnology, Graduate School, University of Gadjah Mada, Yogyakarta 55281, Indonesia
3 Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Gadjah Mada, Yogyakarta, 55281, Indonesia
4 Department of Fisheries, Faculty of Agriculture, Gadjah Mada University, Yogyakarta, 55281, Indonesia
5 Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Gadjah Mada, Yogyakarta, 55281, Indonesia
6 Department of Medical Laboratory Technology, STIKES Karya Putra Bangsa, Tulungagung, East Java, 66291, Indonesia
7 Eijkman Research Center for Molecular Biology, National Research and Innovation Agency, Meatpro Building, Eijkman Research Center for Molecular Biology, National Research and Innovation Agency, Bogor, West Java, 16911, Indonesia
8 Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Gadjah Mada, Yogyakarta, 55281, Indonesia
doi
10.48309/chemm.2026.573435.2096چکیده
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 309-amino-acid construct incorporating the 50S ribosomal protein L7/L12 as a TLR4-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 TLR4 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-28b (+) expression system. Recombinant protein expression in Escherichia coli BL21 (DE3) was validated by molecular analysis and SDS–PAGE, revealing a protein of approximately 35.17 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.
