Plant N-glycan profiling of minute amounts of material - Normandie Université Access content directly
Journal Articles Analytical Biochemistry Year : 2008

Plant N-glycan profiling of minute amounts of material


Development of convenient strategies for identification of plant N-glycan profiles has been driven by the emergence of plants as an expression system for therapeutic proteins. In this article, we reinvestigated qualitative and quantitative aspects of plant N-glycan profiling. The extraction of plant proteins through a phenol/ammonium acetate procedure followed by deglycosylation with peptide N-glycosidase A (PNGase A) and coupling to 2-aminobenzamide provides an oligosaccharide preparation containing reduced amounts of contaminants from plant cell wall polysaccharides. Such a preparation was also suitable for accurate qualitative and quantitative evaluation of the N-glycan content by mass spectrometry. Combining these approaches allows the profiling to be carried out from as low as 500 mg of fresh leaf material. We also demonstrated that collision-induced dissociation (CID) mass spectrometry in negative mode of N-glycans harboring alpha(1,3)- or alpha(1,6)-fucose residue on the proximal GlcNAc leads to specific fragmentation patterns, thereby allowing the discrimination of plant N-glycans from those arising from mammalian contamination.

Dates and versions

hal-01974002 , version 1 (08-01-2019)



Martial Séveno, Gleysin Cabrera, Ada Triguero, Carole Burel, Jérôme Leprince, et al.. Plant N-glycan profiling of minute amounts of material. Analytical Biochemistry, 2008, 379 (1), pp.66-72. ⟨10.1016/j.ab.2008.04.034⟩. ⟨hal-01974002⟩
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