Recombinant influenza virus-like particles (vlps) produced in transgenic plants

Abstract

A method for synthesizing influenza virus-like particles (VLPs) within a plant or a portion of a plant is provided. The method involves expression of influenza HA in plants and the purification by size exclusion chromatography. The invention is also directed towards a VLP comprising influenza HA protein and plant lipids. The invention is also directed to a nucleic acid encoding influenza HA as well as vectors. The VLPs may be used to formulate influenza vaccines, or may be used to enrich existing vaccines.

1 - 18 . (canceled) 
     
     
         19 . A plant or plant cell comprising influenza virus like particle (VLP). 
     
     
         20 . The plant or plant cell of  claim 19 , wherein the VLP comprise influenza virus hemagglutinin (HA) protein and one or more than one lipid derived from the plant or plant cell. 
     
     
         21 . The plant or plant cell of  claim 20 , wherein the HA is a type A influenza, a type B influenza, or is a subtype of type A influenza, selected from the group consisting of H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15 and H16. 
     
     
         22 . The plant or plant cell of  claim 20 , wherein the influenza hemagglutinin (HA) has 70%-100% sequence identity with the amino acid sequence of SEQ ID NO: 9, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 74, SEQ ID NO: 75 or SEQ ID NO: 76. 
     
     
         23 . The plant or plant cell of  claim 20 , wherein the HA protein comprises one or more than one plant-specific N-glycans, or modified N-glycans. 
     
     
         24 . The plant or plant cell of  claim 20 , wherein the plant or plant cell further comprises a nucleic acid comprising a nucleotide sequence encoding the influenza hemagglutinin (HA), the HA being operatively linked to a regulatory element that is operative in the plant or plant cell. 
     
     
         25 . A plant extract derived from the plant or plant cell of  claim 19 . 
     
     
         26 . A composition comprising the plant extract of  claim 25  and a pharmaceutical acceptable carrier. 
     
     
         27 . A method of producing a plant extract from a plant or plant cell comprising influenza virus like particle (VLP), the method comprising:
 a) providing the plant or plant cell of  claim 19 ;   b) harvesting the plant or plant cell;   c) producing a plant extract from the harvested plant or plant cell, wherein the plant extract comprises influenza VLP.   
     
     
         28 . A plant extract produced by the method of  claim 27 . 
     
     
         29 . A composition comprising the plant extract of  claim 28  and a pharmaceutical acceptable carrier. 
     
     
         30 . A plant or plant cell comprising a nucleic acid comprising a nucleotide sequence encoding an influenza hemagglutinin (HA), the HA being operatively linked to a regulatory element that is operative in the plant or plant cell. 
     
     
         31 . The plant or plant cell of  claim 30 , wherein the plant or plant cell further comprises influenza virus like particle (VLP). 
     
     
         32 . The plant or plant cell of  claim 31 , wherein the VLP comprise influenza virus hemagglutinin (HA) protein and one or more than one lipid derived from the plant or plant cell. 
     
     
         33 . A plant extract derived from the plant or plant cell of  claim 30 . 
     
     
         34 . A composition comprising the plant extract of  claim 33  and a pharmaceutical acceptable carrier. 
     
     
         35 . A method of producing a plant extract from a plant or plant cell comprising influenza virus like particle (VLP), the method comprising:
 a) providing the plant or plant cell of  claim 30 ;   b) harvesting the plant or plant cell;   c) producing a plant extract from the harvested plant or plant cell, wherein the plant extract comprises influenza VLP.   
     
     
         36 . A plant extract produced by the method of  claim 35 . 
     
     
         37 . A composition comprising the plant extract of  claim 36  and a pharmaceutical acceptable carrier. 
     
     
         38 . A plant or plant cell comprising influenza hemagglutinin (HA).