Designing a multi-epitope vaccine against African swine fever virus using immunoinformatics approach.

African swine fever (ASF) is a highly contagious and fatal haemorrhagic disease affecting domestic and wild pigs, with no effective vaccine currently available. The lack of an effective vaccine has hindered global ASF control efforts, leading to devastating economic losses in the swine industry. Traditional vaccine development approaches have faced challenges due to ASFV's genetic complexity and immune evasion strategies. Therefore, this study aims to leverage immunoinformatic approaches to facilitate acceleration of the early stages of vaccine development, optimizing resource utilization and time efficiency while providing a rational design for a potent multi-epitope vaccine against ASFV. In this study, a multi-epitope vaccine against ASFV was designed using an in-silico approach incorporating epitopes from conserved ASFV genes - B646L (p72), CP204L (p30), E183L (p54) and EP402R (CD2v). Promising epitopes that were antigenic and non-allergenic were used for the vaccine construct along with suitable adjuvant and linkers. Further analyses of the construct interpreted the physico-chemical properties, secondary and tertiary structure prediction and validation. The docking and molecular dynamics analysis of the docked complex (vaccine construct and SLA-1 0401) were performed. The docking analysis demonstrated that the vaccine construct binds well with SLA-1 0401 and the molecular dynamics analysis confirmed its strong binding affinity. The vaccine construct was confirmed as stable through normal mode analysis (NMA). Immune simulations demonstrated that this multi-epitope vaccine construct generates a strong adaptive immune response including both humoral and cell-mediated immunity. The sequence of the vaccine construct was further codon optimized with better CAI and GC content, for enhanced expression in the host Sus scrofa. Finally, the optimized sequence of vaccine construct was cloned into the plasmid pVAX1-eGFP. These in-silico results prove that the designed multi-epitope vaccine is potentially effective and warrants for further in vitro and in vivo studies to confirm the efficiency of the vaccine against ASFV.