P40-S Isobaric Labeling for Comparative Glycomics

A novel strategy, based on the use of isobaric labeling, is introduced for quantitative glycomics. In this approach, glycans are permethylated with either (13)CH(3)I or (12)CH(2)DI. This pair of reagents has the same nominal mass but differ in their exact mass by 0.002922 Da per label. This mass dif...

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Main Authors: Orlando, R., Atwood, J. A., Cheng, L., York, W. S.
Formato: Artigo
Idioma:English
Publicado em: The Association of Biomolecular Resource Facilities 2007
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Poster Abstracts: Carbohydrate Analysis
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC2292021/
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spelling pubmed-22920212008-05-27 P40-S Isobaric Labeling for Comparative Glycomics Orlando, R. Atwood, J. A. Cheng, L. York, W. S. J Biomol Tech Poster Abstracts: Carbohydrate Analysis A novel strategy, based on the use of isobaric labeling, is introduced for quantitative glycomics. In this approach, glycans are permethylated with either (13)CH(3)I or (12)CH(2)DI. This pair of reagents has the same nominal mass but differ in their exact mass by 0.002922 Da per label. This mass difference is difficult to resolve with current mass spectrometers in cases where only a single label is attached to the analyte. However, glycans typically contain multiple hydroxyl groups, which increase the mass difference between the two samples. Since the number of hydroxyl groups increases with the mass of the glycan, the difference between these isobaric species also increases and thus the resolution needed is approximately 25,000 ΔM/M and is independent of the glycan’s size for typical N- and O-linked species. The advantages of this labeling strategy are numerous, and result primarily from the isobaric ions appearing at the same nominal mass-to-charge ratio. This characteristic leads to increased ion intensity as ions from both samples are not distributed between isotopic species having different m/z values. The small mass difference between these isobars allows the two species to be simultaneously selected for MS(n) analysis, permitting the relative quantitation of isomeric glycans. This procedure was successfully used to analyze N- and O-linked oligosaccharides released from a standard glycoprotein and from human serum. This strategy is directly applicable to oligosaccharides from other sources, such as glycolipds. The concept of isobaric labeling is expected to be applicable to other types of “omics” analyses with other derivatizing agents. The Association of Biomolecular Resource Facilities 2007-02 /pmc/articles/PMC2292021/ Text en Copyright © Copyright 2007 by The Association of Biomolecular Resource Facilities
institution US National Library of Medicine
collection PubMed Central
language English
format Article
topic Poster Abstracts: Carbohydrate Analysis
spellingShingle Poster Abstracts: Carbohydrate Analysis
Orlando, R.
Atwood, J. A.
Cheng, L.
York, W. S.
P40-S Isobaric Labeling for Comparative Glycomics
description A novel strategy, based on the use of isobaric labeling, is introduced for quantitative glycomics. In this approach, glycans are permethylated with either (13)CH(3)I or (12)CH(2)DI. This pair of reagents has the same nominal mass but differ in their exact mass by 0.002922 Da per label. This mass difference is difficult to resolve with current mass spectrometers in cases where only a single label is attached to the analyte. However, glycans typically contain multiple hydroxyl groups, which increase the mass difference between the two samples. Since the number of hydroxyl groups increases with the mass of the glycan, the difference between these isobaric species also increases and thus the resolution needed is approximately 25,000 ΔM/M and is independent of the glycan’s size for typical N- and O-linked species. The advantages of this labeling strategy are numerous, and result primarily from the isobaric ions appearing at the same nominal mass-to-charge ratio. This characteristic leads to increased ion intensity as ions from both samples are not distributed between isotopic species having different m/z values. The small mass difference between these isobars allows the two species to be simultaneously selected for MS(n) analysis, permitting the relative quantitation of isomeric glycans. This procedure was successfully used to analyze N- and O-linked oligosaccharides released from a standard glycoprotein and from human serum. This strategy is directly applicable to oligosaccharides from other sources, such as glycolipds. The concept of isobaric labeling is expected to be applicable to other types of “omics” analyses with other derivatizing agents.
author Orlando, R.
Atwood, J. A.
Cheng, L.
York, W. S.
author_facet Orlando, R.
Atwood, J. A.
Cheng, L.
York, W. S.
author_sort Orlando, R.
title P40-S Isobaric Labeling for Comparative Glycomics
title_short P40-S Isobaric Labeling for Comparative Glycomics
title_full P40-S Isobaric Labeling for Comparative Glycomics
title_fullStr P40-S Isobaric Labeling for Comparative Glycomics
title_full_unstemmed P40-S Isobaric Labeling for Comparative Glycomics
title_sort p40-s isobaric labeling for comparative glycomics
publisher The Association of Biomolecular Resource Facilities
publisher_facet The Association of Biomolecular Resource Facilities
publishDate 2007
url https://ncbi.nlm.nih.gov/pmc/articles/PMC2292021/
_version_ 1760534654345019392

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