High-throughput quantitative analysis of plant N-glycan using a DNA sequencer

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Title
High-throughput quantitative analysis of plant N-glycan using a DNA sequencer
Author(s)
Kyung Jin Lee; J H Jung; Jung Mi Lee; Y So; Oh Suk Kwon; N Callewaert; H A Kang; K Ko; Doo-Byoung Oh
Bibliographic Citation
Biochemical and Biophysical Research Communications, vol. 380, no. 2, pp. 223-229
Publication Year
2009
Abstract
High-throughput quantitative analytical method for plant N-glycan has been developed. All steps, including peptide N-glycosidase (PNGase) A treatment, glycan preparation, and exoglycosidase digestion, were optimized for high-throughput applications using 96-well format procedures and automatic analysis on a DNA sequencer. The glycans of horseradish peroxidase with plant-specific core α(1,3)-fucose can be distinguished by the comparison of the glycan profiles obtained via PNGase A and F treatments. The peaks of the glycans with (91%) and without (1.2%) α(1,3)-fucose could be readily quantified and shown to harbor bisecting β(1,2)-xylose via simultaneous treatment with α(1,3)-mannosidase and β(1,2)-xylosidase. This optimized method was successfully applied to analyze N-glycans of plant-expressed recombinant antibody, which was engineered to contain a minor amount of glycan harboring β(1,2)-xylose. These results indicate that our DNA sequencer-based method provides quantitative information for plant-specific N-glycan analysis in a high-throughput manner, which has not previously been achieved by glycan profiling based on mass spectrometry.
Keyword
α(1,3)-Fucoseβ(1,2)-XyloseDNA sequencerGlycan analysisPlant N-glycan
ISSN
0006-291X
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.bbrc.2009.01.070
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
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