Uncovering the Unique Epitopes: Validation of In-Silico Synthesized Tetravalent Dengue Vaccine (dvac) Through Cloning, Expression, and Immunogenic Analysis.

Dengue is an arthropod borne viral illness which infects millions of people each year. Despite the huge global losses, an effective medication is still unavailable against the disease. For vaccine development, reverse vaccinology approach is being tremendously used as it saves time, energy, cost and resources by selecting the potential epitopes for developing an effective vaccine formulation. In our previous study, we have also used reverse vaccinology approach to propose a multi-epitope vaccine formulation (dvac) against dengue, by targeting the conserved epitopes from all of its serotypes. In this study, we have performed population coverage analysis and molecular dynamic simulation studies which predicted that dvac could offer protection to 98.5% of world population against dengue and forms stable interactions with TLR3/TLR4 receptors, respectively. Moreover, we have performed in-lab gene cloning and expression analysis of dvac in Escherichia coli. The purified dvac is subjected to anti-NS1, anti-dNS1 and anti-EDIII antibodies, raised in rabbits, and the ELISA results confirmed the individuality and intactness of the epitopes incorporated in the refolded dvac molecule. The results from this study endorse the use of dvac as a potential dengue vaccine candidate and also support the reverse vaccinology approach for developing novel therapeutic formulations.