A method of producing Enterovirus-like particles (EVLPs) in a plant, portion of the plant, or plant cell comprising: i-a) introducing by transformation into the plant, portion of the plant, or plant cell a first nucleic acid comprising a first regulatory region active in the plant operatively linked to a first nucleotide sequence encoding an Enterovirus polyprotein, wherein the Enterovirus polyprotein consist of Enterovirus 71 polyprotein PI, the first regulatory region comprising a first promoter operatively linked to a first enhancer; introducing by transformation into the plant, portion of the plant, or plant cell a second nucleic acid comprising a second regulatory region active in the plant and operatively linked to a second nucleotide sequence encoding an Enterovirus 71 3CD protease, the second regulatory region comprising a second promoter operatively linked to a second enhancer; or i-b) providing the plant, portion of the plant or plant cell comprising a first nucleic acid comprising a first regulatory region active in the plant operatively linked to a first nucleotide sequence encoding an Enterovirus polyprotein, wherein the Enterovirus polyprotein consist of Enterovirus 71 polyprotein PI, the first regulatory region comprising a first promoter operatively linked to a first enhancer; and a second nucleic acid comprising a second regulatory region active in the plant operatively linked to a second nucleotide sequence encoding an Enterovirus 71 3CD protease, the second regulatory region comprising a second promoter operatively linked to a second enhancer; ii) incubating the plant, portion of the plant, or plant cell under conditions that permit the expression of the first and second nucleic acid, thereby producing the EVLPs; and optionally iii) harvesting the plant, portion of the plant, or plant cell and purifying the EVLPs. i-a) introducing by transformation into the plant, portion of the plant, or plant cell a first nucleic acid comprising a first regulatory region active in the plant operatively linked to a first nucleotide sequence encoding an Enterovirus polyprotein, wherein the Enterovirus polyprotein consist of Enterovirus 71 polyprotein PI, the first regulatory region comprising a first promoter operatively linked to a first enhancer; introducing by transformation into the plant, portion of the plant, or plant cell a second nucleic acid comprising a second regulatory region active in the plant and operatively linked to a second nucleotide sequence encoding an Enterovirus 71 3CD protease, the second regulatory region comprising a second promoter operatively linked to a second enhancer; or i-b) providing the plant, portion of the plant or plant cell comprising a first nucleic acid comprising a first regulatory region active in the plant operatively linked to a first nucleotide sequence encoding an Enterovirus polyprotein, wherein the Enterovirus polyprotein consist of Enterovirus 71 polyprotein PI, the first regulatory region comprising a first promoter operatively linked to a first enhancer; and a second nucleic acid comprising a second regulatory region active in the plant operatively linked to a second nucleotide sequence encoding an Enterovirus 71 3CD protease, the second regulatory region comprising a second promoter operatively linked to a second enhancer; ii) incubating the plant, portion of the plant, or plant cell under conditions that permit the expression of the first and second nucleic acid, thereby producing the EVLPs; and optionally iii) harvesting the plant, portion of the plant, or plant cell and purifying the EVLPs.