A zoonotic Orthopoxvirus called Monkeypox Virus (MPXV) causes smallpox-like infections that affect people. Worldwide, the World Health Organization, abbreviated as WHO has been recorded to be growing in quantity of monkeypox (MPX) cases since May 2022.Currently, there are lots of problems with utilizing the smallpox vaccination to prevent monkeypox, and the treatment of infections with the disease is not clinically established. Considering this necessity and the rising number of cases of treatment resistance, creating a more potent and improved vaccination against the monkeypox virus is extremely desirable. In the present study, reverse vaccinology and several other bioinformatics and immunoinformatics tools were utilized to design multi-epitopes-based vaccine against MPXV by exploring four probable cell proteins like E8L, A28, COP-A44L and COP-B7R. The potential epitopes T-cell and B-cell were predicted from the proteins and connected with the Support of adjuvants and linkers. The predicted epitopes' physiochemical properties were evaluated, and only probable antigenic, non-allergic, and non-toxic epitopes were utilized in the multi-epitope vaccine design. The 3D structure of the designed vaccine is predicted, refined and validated for molecular docking with human immune receptor TLR4 demonstrated increased binding interaction. The designed vaccine construct was reverse transcribed and modified for E. coli strain K12 preceding inclusion inside pET28a (+) vector for its heterologous cloning and expression. Immunological responses were found to be enhanced by the IMMSIM server. In conclusion, multi-epitope vaccine candidates were created and their effectiveness has been verified. The strategy developed in this study could hold considerable importance effects on the early detection and therapy of infectious diseases brought on by the monkeypox virus.