Improved sample preparation for CE-LIF analysis of plant N-glycans

被引:6
|
作者
Nagels, Bieke [1 ,2 ]
Santens, Francis [3 ]
Weterings, Koen [2 ]
Van Damme, Els J. M. [1 ]
Callewaert, Nico [3 ,4 ]
机构
[1] Univ Ghent, Dept Mol Biotechnol, Lab Biochem & Glycobiol, Fac Biosci Engn, B-9000 Ghent, Belgium
[2] Bayer BioSci NV, Ghent, Belgium
[3] VIB, Dept Mol Biomed Res, Unit Med Biotechnol, Ghent, Belgium
[4] Univ Ghent, Fac Sci, Dept Biochem & Microbiol, B-9000 Ghent, Belgium
关键词
Arabidopsis thaliana; CE-LIF; In-gel release; N-glycan analysis; SDS-PAGE; NICOTIANA-BENTHAMIANA; PROCESSING ENZYMES; GLYCOSYLATION; GLYCOPROTEINS; PROTEINS; ANTIBODIES; CELLS; EXPRESSION; GENERATION; GROWTH;
D O I
10.1002/elps.201100268
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In view of glycomics studies in plants, it is important to have sensitive tools that allow one to analyze and characterize the N-glycans present on plant proteins in different species. Earlier methods combined plant-based sample preparations with CE-LIF N-glycan analysis but suffered from background contaminations, often resulting in non-reproducible results. This publication describes a reproducible and sensitive protocol for the preparation and analysis of plant N-glycans, based on a combination of the in-gel release method and N-glycan analysis on a multicapillary DNA sequencer. Our protocol makes it possible to analyze plant N-glycans starting from low amounts of plant material with highly reproducible results. The developed protocol was validated for different plant species and plant cells.
引用
收藏
页码:3482 / 3490
页数:9
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