Background:　Enterovirus　71　(EV71)　is　the　most　common　causative　pathogens　of　hand,　foot　and　mouth　disease　(HFMD)　associated　with　severe　neurological　complications.　There　is　a　great　need　to　develop　prophylactic　vaccine　against　EV71　infection.　Results:　EV71　virus-like　particle　(VLP)　was　produced　in　yeast　expression　system　by　the　co-expression　of　four　EV71　structural　proteins　VP1-VP4.　Immunization　with　the　recombinant　VLPs　elicited　potent　anti-EV71　antibody　responses　in　adult　mice　and　anti-VLP　sera　were　able　to　neutralize　EV71　virus　in　vitro.　Neonatal　mice　model　demonstrated　VLP　immunization　conferred　protection　to　suckling　mice　against　the　lethal　viral　challenge.　Conclusions:　Co-expression　of　four　EV71　structural　proteins　VP1-VP4　in　yeast　expression　systems　is　an　effective　method　to　produce　EV71　VLPs.　VLP-based　vaccine　shows　great　potential　to　prevent　EV71　infection.　©　2016　Wang　et　al.