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Plant Biotechnol J. 2015 Jun;13(5):717-25. doi: 10.1111/pbi.12301. Epub 2014 Dec 18.

Biochemical composition of haemagglutinin-based influenza virus-like particle vaccine produced by transient expression in tobacco plants.

Author information

1
Laboratoire Glyco-MEV EA 4358, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.
2
Medicago inc., Québec, QC, Canada.
3
PISSARO Proteomic Platform, Institut de Recherche et d'Innovation Biomédicale, Normandie Université, Mont-Saint-Aignan, France.

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.

KEYWORDS:

N-glycosylation; haemagglutinin; lipid raft; plant; virus-like particles

PMID:
25523794
DOI:
10.1111/pbi.12301
[Indexed for MEDLINE]
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