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Proceedings Paper

Glycoprotein analysis using enzymatic digestion and MALDI-TOF MS
Author(s): Rich Kornfeld; James W. Kenny; Scot R. Weinberger; Yi Yang; Ron Orlando
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Paper Abstract

A sensitive and facile method is described to identify the glycosylation sites and site-specific heterogeneity in the carbohydrate portion of glycoproteins. In this procedure, the peptide backbone of the glycoprotein is cleaved enzymatically. The peptide/glycopeptide mixture is divided into three fractions. The first fraction is analyzed directly by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), while the other two aliquots are analyzed by MALDI-TOF MS after enzymatic release of the N-linked and N- and O-linked chains. Comparison of these mass spectra provides the molecular weight of each carbohydrate side chain and of the peptide to which it is attached. This information combined with the protein's amino acid sequence identifies the glycosylation sites and provides information concerning site-specific oligosaccharide heterogeneity. This approach is faster and simpler than procedures currently used for glycosylation site mapping and can be performed on as little as 10 picomoles of glycoprotein.

Paper Details

Date Published: 1 April 1996
PDF: 9 pages
Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); doi: 10.1117/12.237629
Show Author Affiliations
Rich Kornfeld, Hewlett-Packard Co. (United States)
James W. Kenny, Hewlett-Packard Co. (United States)
Scot R. Weinberger, Hewlett-Packard Co. (United States)
Yi Yang, Univ. of Georgia (United States)
Ron Orlando, Univ. of Georgia (United States)

Published in SPIE Proceedings Vol. 2680:
Ultrasensitive Biochemical Diagnostics
Gerald E. Cohn; Steven A. Soper; C. H. Winston Chen, Editor(s)

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