Small-Scale Enzymatic Digestion of Glycoproteins and Proteoglycans for Analysis of Oligosaccharides by LC-MS and FACE Gel Electrophoresis View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2008-02-20

AUTHORS

Ruby P. Estrella , John M. Whitelock , Rebecca H. Roubin , Nicolle H. Packer , Niclas G. Karlsson

ABSTRACT

Structural characterization of oligosaccharides from proteoglycans and other glycoproteins is greatly enhanced through the use of mass spectrometry and gel electrophoresis. Sample preparation for these sensitive techniques often requires enzymatic treatments to produce oligosaccharide sequences for subsequent analysis. This chapter describes several small-scale methods for in-gel, on-blot, and in-solution enzymatic digestions in preparation for graphitized carbon liquid chromatography-mass spectrometry (LC-MS) analysis, with specific applications indicated for glycosaminoglycans (GAGs) and N-linked oligosaccharides. In addition, accompanying procedures for oligosaccharide reduction by sodium borohydride, sample desalting via carbon microcolumn, desialylation by sialidase enzyme treatment, and small-scale oligosaccharide species fractionation are included. Fluorophore-assisted carbohydrate electrophoresis (FACE) is another useful method to isolate derivatized oligosaccharides. Overall, the modularity of these techniques provides ease and flexibility for use in conjunction with mass spectrometric and electrophoretic tools for glycomic research studies. More... »

PAGES

171-192

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-59745-022-5_13

DOI

http://dx.doi.org/10.1007/978-1-59745-022-5_13

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1040140949

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/19277552


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