Glycan reductive isotope labeling for quantitative glycomics

Many diseases and disorders are characterized by quantitative and/or qualitative changes in complex carbohydrates. Mass spectrometry methods show promise in monitoring and detecting these important biological changes. Here we report a new glycomics method, termed glycan reductive isotope labeling (G...

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Bibliographic Details
Published in:Analytical biochemistry 2009-04, Vol.387 (2), p.162-170
Main Authors: Xia, Baoyun, Feasley, Christa L., Sachdev, Goverdhan P., Smith, David F., Cummings, Richard D.
Format: Article
Language:English
Subjects:
Animals
Glycans
Glycomics - methods
Glycoproteins
Glycoproteins - blood
Glycoproteins - chemistry
Humans
Isotope Labeling
MALDI
Mass spectrometry
Mice
Oligosaccharides
Polysaccharides - analysis
Polysaccharides - blood
Polysaccharides - chemistry
Reductive amination
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Summary:Many diseases and disorders are characterized by quantitative and/or qualitative changes in complex carbohydrates. Mass spectrometry methods show promise in monitoring and detecting these important biological changes. Here we report a new glycomics method, termed glycan reductive isotope labeling (GRIL), where free glycans are derivatized by reductive amination with the differentially coded stable isotope tags [ 12C 6]aniline and [ 13C 6]aniline. These dual-labeled aniline-tagged glycans can be recovered by reverse-phase chromatography and can be quantified based on ultraviolet (UV) absorbance and relative ion abundances. Unlike previously reported isotopically coded reagents for glycans, GRIL does not contain deuterium, which can be chromatographically resolved. Our method shows no chromatographic resolution of differentially labeled glycans. Mixtures of differentially tagged glycans can be directly compared and quantified using mass spectrometric techniques. We demonstrate the use of GRIL to determine relative differences in glycan amount and composition. We analyze free glycans and glycans enzymatically or chemically released from a variety of standard glycoproteins, as well as human and mouse serum glycoproteins, using this method. This technique allows linear relative quantitation of glycans over a 10-fold concentration range and can accurately quantify sub-picomole levels of released glycans, providing a needed advancement in the field of glycomics.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2009.01.028