Picornavirus-like particle production in plants

A plant extract comprising an Enterovirus-like particles (EVLPs) produced by a method comprising:

• a) introducing into a plant, portion of a plant, or a plant cell a first nucleic acid comprising a first regulatory region active in the plant operatively linked to a nucleotide sequence encoding an Enterovirus polyprotein, wherein the Enterovirus polyprotein consists of Enterovirus 71 polyprotein P1;
• b) introducing a second nucleic acid comprising a second regulatory region active in the plant and operatively linked to a second nucleotide sequence encoding one or more Enterovirus 71 3C or 3CD protease into the plant, portion of the plant, or plant cell;
• c) incubating the plant, portion of the plant, or plant cell under conditions that permit expression of the first and second nucleic acid to produce the Enterovirus 71 polyprotein P1 and the one or more Enterovirus 71 3C or 3CD protease, the Enterovirus 71 polyprotein P1 being processed into structural proteins VP1, VP3, and VP0 or VP1, VP2, VP3 and VP4, thereby producing the EVLP;
• and
• d) harvesting the plant, portion of the plant, or plant cell to produce the plant extract.

A plant extract comprising an Enterovirus-like particle (EVLP) produced by a method comprising:

• a) 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 P1 and a second nucleic acid comprising a second regulatory region active in the plant operatively linked to a second nucleotide sequence encoding one or more Enterovirus 71 3C or 3CD protease;
• b) incubating the plant, portion of the plant or plant cell under conditions that permit expression of the nucleic acids to produce the Enterovirus 71 polyprotein P1 and the one or more Enterovirus 71 3C or 3CD protease, the Enterovirus 71 polyprotein P1 being processed into structural proteins VP1, VP3, and VP0 or VP1, VP2, VP3 and VP4, thereby producing the EVLP;
• and
• c) harvesting the plant, portion of the plant, or plant cell to produce the plant extract.