High-throughput quantitative analysis of plant N-glycan using a DNA sequencer
- Authors
- Lee, Kyung Jin; Jung, Jin-Hee; Lee, Jung Mi; So, Yangkang; Kwon, Ohsuk; Callewaert, Nico; Kang, Hyun Ah; Ko, Kisung; Oh, Doo-Byoung
- Issue Date
- Mar-2009
- Publisher
- ACADEMIC PRESS INC ELSEVIER SCIENCE
- Keywords
- Plant N-glycan; DNA sequencer; Glycan analysis; alpha(1,3)-Fucose; beta(1,2)-Xylose
- Citation
- BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, v.380, no.2, pp 223 - 229
- Pages
- 7
- Journal Title
- BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
- Volume
- 380
- Number
- 2
- Start Page
- 223
- End Page
- 229
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/23273
- DOI
- 10.1016/j.bbrc.2009.01.070
- ISSN
- 0006-291X
1090-2104
- 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 alpha(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%) alpha(1,3)-fucose could be readily quantified and shown to harbor bisecting beta(1,2)-xylose via simultaneous treatment with alpha(1,3)-mannosidase and beta(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 P(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. (c) 2009 Elsevier Inc. All rights reserved.
- Files in This Item
- There are no files associated with this item.
- Appears in
Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.