ABSTRACTCurrent vaccines used for polio are live attenuated oral polio vaccine and inactivated polio vaccine. Recently, in India, the relapse of virulence was observed in attenuated viruses resulting in catastrophic effects. Therefore, the need for the development of multi-epitope subunit vaccine was realised and an immunoinformatics approach to design a multi-epitope subunit vaccine was conceived. Capsid proteins of all the three types of polio strains were utilised to predict major histocompatibility complex class-1 as well as class-2 epitopes. The subunit vaccine was designed with β-defensin at N-terminal followed by cytotoxic T-lymphocytes epitopes and helper T-lymphocytes epitopes connected by compatible linkers. The vaccine construct was further modelled and docked against TLR4 receptor. The high affinity of the construct towards the receptor was observed in the docking study and also substantiated by a 20 ns simulation of the complex. The vaccine construct was cloned in-silico for expression of the protein effectively in a prokaryotic system (Escherichia coli strain K12). Immuno-simulation of the construct was found to elicit immunoglobulin production effectively in the human body. This designed multi-epitope subunit vaccine is capable of immune response and further studies will help us understand the feasibility of this multi-epitope subunit vaccine.