Abstract: Described herein is a method for the production of flavivirus virus-like particles (VLPs) in a yeast system. In some cases, flavivirus structural proteins are expressed in a Pichia pastoris strain and isolated using pressurized mechanical lysis. The disclosed VLPs can be used as immunogenic compositions for the prevention or treatment of flavivirus infection. Also described are isolated nucleic acid molecules encoding a flavivirus capsid (C) protein; a flavivirus premembrane (prM) protein; and at least one flavivirus envelope (E) protein or a flavivirus E protein lacking the transmembrane domain (E?TM), wherein the nucleic acid molecule further encodes at least one foot and mouth disease virus (FMDV) 2A autocatalytic site (2A) positioned between two flavivirus protein coding sequences. Expression of such nucleic acid molecules in a host cell produces flavivirus VLPs.