Medicago Inc.

Centre National De La Recherche Scientifique

Universitè Laval

Université´de Rouen

Plant Bioscience Limited

Agriculture and Agri-Food Canada

Medicago Inc. of Quebec, Canada

Patrice Lerouge

Thomas Paccalet

Loiec Faye

Stephanie Aquin

Muriel Bardor

Veronique Gomord

Louis-Philippe Vezina

Marc-Andre D'aoust

Manon Couture

Marc-Andre D'Aoust

Pierre-Olivier Lavoie

Nathalie Landry

Michele Dargis

Pierre Lavallee

Sonia Trepanier

Laurence Cambron-Fortin

Rejean Desgagnes

Pierre Talbot

Frederic Ors

Dany Paquet

Véronique Gomord

Luc Laurin

Nicole Bechtold

Marc-André D'Aoust

Stéphanie Aquin

Loïc Faye

Loic Faye

Christophe Rihouey

Lucie Poulin

George Peter Lomonossoff

Dominique Michaud

Carole Burel

Frank Sainsbury

Philippe Varennes-Jutras

Ronald E Simard

Vital Hagenimana

Carmen Canizares

Li Liu

Michéle Dargis

Michèle Dargis

Frédéric Ors

Normand Dubuc

Estelle Marquet-Blouin

George Lomonossoff

Christophe Sourrouille

Marc-André D'AOUST

Methods of preparing plant-derived proteins or suprastructure proteins, are provided. The method may comprise obtaining a plant, or plant matter comprising apoplast-localized proteins, or suprastructure proteins, producing a protoplast/spheroplast fraction and apoplast fraction from the plant or plant matter, and recovering the apoplast fraction. The apoplast fraction comprises plant-derived proteins or suprastructure proteins. Alternatively, the proteins, or suprastructure proteins, may be obtained from plant or plant matter comprising plant-derived proteins or suprastructure proteins, by digesting the plant matter using a cell wall degrading enzyme composition to produced a digested fraction. The digested fraction is filtered to produced a filtered fraction, and the plant-derived proteins or suprastructure proteins, are recovered from the filtered fraction.

Method of preparing plant-derived proteins

Methods of preparing plant-derived virus like particles (VLPs) are provided. The method may comprise obtaining a plant, or plant matter comprising apoplast-localized VLPs, producing a protoplast/spheroplast fraction and apoplast fraction from the plant or plant matter, and recovering the apoplast fraction. The apoplast fraction comprises plant-derived VLPs. Alternatively, VLPs may be obtained from plant or plant matter comprising plant-derived VLPs by digesting the plant matter using a cell wall degrading enzyme composition to produced a digested fraction. The digested fraction is filtered to produced a filtered fraction, and the plant-derived VLPs are recovered from the filtered fraction.

Method of preparing plant-derived VLPs

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.

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

A method of increasing the yield, stability, or both of an acid sensitive protein in a plant is provided. The method comprises introducing a first nucleic acid and a second nucleic acid into the plant, or portion of the plant. The first nucleic acid comprises a first regulatory region active in the plant and operatively linked to a nucleotide sequence encoding the acid sensitive protein. The second nucleic acid comprises a second regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a channel protein, for example but not limited to a proton channel protein. The plant or portion of the plant is incubated under conditions that permit the expression of the nucleic acids, thereby increasing the yield of the acid sensitive protein when compared to the yield of the acid sensitive protein produced in the plant or portion of the plant produced under the same conditions, and in the absence of the proton channel protein.

Increasing protein yield in plants

A method of producing a virus like particle (VLP) in a plant is provided. The method comprises introducing a first nucleic acid and a second nucleic acid into the plant, or portion of the plant. The first nucleic acid comprises a first regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a structural virus protein. The second nucleic acid comprises a second regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a channel protein, for example but not limited to a proton channel protein. The plant or portion of the plant is incubated under conditions that permit the expression of the nucleic acids, thereby producing the VLP.

Increasing virus-like particle yield in plants

A method of producing a virus like particle (VLP) in a plant comprising modified hemagglutinin is provided. The method comprises introducing a nucleic acid comprising a regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a modified influenza hemagglutinin (HA) protein into the plant, or portion of the plant, the modified HA protein comprises a modified proteolytic loop. Followed by incubating the plant or portion of the plant under conditions that permit the expression of the nucleic acids, thereby producing the VLP. The modified proteolytic loop may comprise one or more protease cleavage sites exhibiting reduced or abolished cleavage by a protease. The nucleotide sequence encoding the HA may be selected from the group consisting of B HA, C, H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, and H16. Also described is a virus like particle (VLP) produced by the method, and plants expressing the VLP. The virus like particle (VLP) may comprise plant-specific N-glycans, or modified N-glycans.

Influenza virus-like particle production in plants

A method of producing a picornavirus-like particle (PVLP) in a plant is provided. The method comprises introducing a first nucleic acid and a second nucleic acid into the plant, portion of the plant, or a plant cell. The first nucleic acid comprising a first regulatory region active in the plant operatively linked to a nucleotide sequence encoding a polyprotein. The second nucleic acid comprises a second regulatory region active in the plant and operatively linked to a nucleotide sequence encoding one or more protease. The plant, portion of the plant, or plant cell is incubated under conditions that permit the expression of the nucleic acids, thereby producing the PVLP. A PVLP comprising the polyprotein is also provided.

Picornavirus-like particle production in plants

A plant tray for receiving and retaining one or more plant pots includes a plant-receiving tray for use in a high-throughput plant infiltration process used to produce recombinant proteins. The plant tray described has a base retaining tray with an upper surface defining cavities for receiving the plant pots, and a top plate displaceably mounted on the base retaining tray and having apertures, the apertures defining a channel with the corresponding cavities of the bottom tray when aligned. The top plate is displaceable between a locked operative position wherein the apertures and the corresponding cavities are partially misaligned, thereby locking the plant pots in between the top plate and the base retaining tray.

Plant infiltration tray

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

Chimeric influenza virus-like particles comprising hemagglutinin

Recombinant influenza virus-like particles (VLPs) produced in transgenic plants expressing hemagglutinin

It is provided a plant infiltration device for infiltrating plants with an inoculum and method of using same, the device comprising a frame having one or more tray racks adapted for receiving at least one tray of plants to be infiltrated; an automated manipulation mechanism mounted to the frame and operable to displace the plant trays within the device; a plurality of inoculum reservoirs mounted to the frame and containing one or more inoculum fluids; an infiltration tank mounted to the frame and within which the plant trays are received when disposed in the infiltration position; and a vacuum generating device in communication with the infiltration tank to apply a negative pressure within the infiltration tank, thereby opening pores of the plant leaves immersed in the inoculum within the infiltration tank and causing the inoculum fluid to infiltrate into the plant leaves.

Plant infiltration device

Methods of recovering plant-derived proteins or suprastructure proteins, are provided. The method may comprise obtaining a plant, or plant matter comprising apoplast-localized proteins, or suprastructure proteins, loosening the cell wall to produce a plant or plant matter having a loosened cell wall, thus allowing the release of apoplastic content through the cell wall to produce an apoplastic content fraction from the plant or plant matter, and recovering the apoplastic content fraction. The apoplastic content fraction comprises plant-derived proteins or suprastructure proteins.

Method for recovering plant-derived proteins

A method of synthesizing sialic acid in plants, and plants capable of synthesizing sialic acid is provided. Furthermore, a method of producing sialylated protein in a plant is also provided. The method to synthesize sialic acid comprises providing a plant comprising a nucleotide sequence encoding N-acetyl neuraminic acid (Neu5Ac) synthase or Neu5Ac lyase, and expressing the nucleotide sequence thereby synthesizing sialic acid. The plant may also co-express a nucleotide sequence encoding one or more than one of an epimerase, a CMP-Neu5Ac synthase, a CMP-Neu5Ac transporter and a sialyltransferase.

Synthesis of sialic acid in plants

Growing community of inventors

Neuville, Canada

Louis-Philippe Vezina