title: Biochemical composition of haemagglutinin-based influenza virus-like particle vaccine produced by transient expression in tobacco plants.
abstract: Influenza virus-like particles (VLPs) are noninfectious particles resembling the influenza virus representing a promising vaccine alternative to inactivated influenza virions as antigens. Medicago inc. has developed a plant-based VLP manufacturing platform allowing the large-scale production of GMP-grade influenza VLPs. In this article, we report on the biochemical compositions of these plant-based influenza candidate vaccines, more particularly the characterization of the N-glycan profiles of the viral haemagglutinins H1 and H5 proteins as well as the tobacco-derived lipid content and residual impurities. Mass spectrometry analyses showed that all N-glycosylation sites of the extracellular domain of the recombinant haemagglutinins carry plant-specific complex-type N-glycans having core α(1,3)-fucose, core β(1,2)-xylose epitopes and Lewis(a) extensions. Previous phases I and II clinical studies have demonstrated that no hypersensibility nor induction of IgG or IgE directed against these glycans was observed. In addition, this article showed that the plant-made influenza vaccines are highly pure VLPs preparations while detecting no protein contaminants coming either from Agrobacterium or from the enzymes used for the enzyme-assisted extraction process. In contrast, VLPs contain few host cell proteins and glucosylceramides associated with plant lipid rafts. Identification of such raft markers, together with the type of host cell impurity identified, confirmed that the mechanism of VLP formation in planta is similar to the natural process of influenza virus assembly in mammals.
authors.affiliation: Laboratoire Glyco-MEV EA 4358, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.|Medicago inc., Québec, QC, Canada.
authors.forename: François
authors.initials: F
authors.lastname: Le Mauff
authors.affiliation: Medicago inc., Québec, QC, Canada.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Mercier
authors.affiliation: PISSARO Proteomic Platform, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Chan
authors.affiliation: Laboratoire Glyco-MEV EA 4358, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Burel
authors.affiliation: PISSARO Proteomic Platform, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.
authors.forename: David
authors.initials: D
authors.lastname: Vaudry
authors.affiliation: Laboratoire Glyco-MEV EA 4358, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Bardor
authors.affiliation: Medicago inc., Québec, QC, Canada.
authors.forename: Louis-Philippe
authors.initials: LP
authors.lastname: Vézina
authors.affiliation: Medicago inc., Québec, QC, Canada.
authors.forename: Manon
authors.initials: M
authors.lastname: Couture
authors.affiliation: Laboratoire Glyco-MEV EA 4358, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.
authors.forename: Patrice
authors.initials: P
authors.lastname: Lerouge
authors.affiliation: Medicago inc., Québec, QC, Canada.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Landry
mesh_terms: D000595:Amino Acid Sequence; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D015870:Gene Expression; D006031:Glycosylation; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000276:immunology*; D007251:Influenza, Human / Q000517:prevention & control*; D021962:Membrane Microdomains; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000737:chemistry; D030821:Plants, Genetically Modified; D011134:Polysaccharides / Q000737:chemistry; D011994:Recombinant Proteins; D013107:Sphingolipids / Q000737:chemistry; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D058425:Vaccines, Virus-Like Particle / Q000276:immunology
chemical_list: D000939:Epitopes; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D010743:Phospholipids; D011134:Polysaccharides; D011994:Recombinant Proteins; D013107:Sphingolipids; D058425:Vaccines, Virus-Like Particle
keywords: N-glycosylation; haemagglutinin; lipid raft; plant; virus-like particles