Designing and Cloning of the Gene Vaccine Carrying the Viral Haemorrhagic Septicaemia Multi‐Epitope Gene in the pNZ8121 Secretion Vector

The viral haemorrhagic septicaemia virus (VHSV) is considered a very significant disease affecting fish. The objective of this project is to develop a multi-epitope vaccine targeting viral hemorrhagic fever using the reverse vaccinology approach. Epitope prediction was conducted using the immunogenic components of VHSVV. A multi-epitope vaccine was developed by combining the immunogenic proteins' most potent B- and T-cell epitopes with the adjuvant. Following that, a comprehensive evaluation was carried out on many facets of the formulated vaccine, including its physicochemical properties, antigenic profile, secondary structure and tertiary structure. In addition, the molecular docking methodology was used to investigate the interaction between the suggested vaccination and its Toll-like receptor 4 (TLR-4) receptor. The vaccine's nucleotide sequence was later altered to enhance its production in Lactococcus lactis. The present investigation's results suggest that the produced vaccine demonstrated stability by its molecular weight of 58,769.30 Da and antigenicity score of 0.5283. Furthermore, the analysis of the vaccine's composition revealed that it included 12.43% alpha-helix, with 87.88% of its residues in the desired region. The vaccine under consideration demonstrated successful docking to its TLR-4, yielding the minimum energy value of -4005.09. Subsequently, the optimum complex was identified as -19.38 (kcal/mol). On the basis of the acquired results, the proposed vaccine has promising efficacy in mitigating fish infection resulting from VHSV. Our research indicates that peptide vaccination might be a beneficial option for preventing VHSV.