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In loop insertion approaches, epitopes are introduced into the surface-exposed loops of capsid proteins from VLPs to induce strong neutralizing responses. In genetic procedures, small or entire proteins are accommodated on the VLP surface via loop insertion, N-terminus/C-terminus (N/C-ter) modification, or the mutation of amino acid residues. Finally, different characterization techniques required to examine the size, charge, aggregation, and composition of VLPs are described. Moreover, the structural classification, morphology, and methods to functionalize the surface of VLPs are discussed. We consulted different databases to present a general scenario about viruses and how VLPs are produced in eukaryotic and prokaryotic cell lines to entrap therapeutic cargo. This review compiles the recent scientific literature about the fundamentals of VLPs with biomedical applications. However, depending upon the intrinsic features of the native virus from which they are produced, the therapeutic performance of VLPs can vary. Virus-like particles (VLPs) are self-assembled nanoparticles whose intrinsic properties, such as heterogeneity, and highly ordered structural organization are exploited to prepare vaccines imaging agents construct nanobioreactors cancer treatment approaches or deliver drugs, genes, and enzymes. Nanotechnology is a fast-evolving field focused on fabricating nanoscale objects for industrial, cosmetic, and therapeutic applications.