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A Quantitative Glycomics and Proteomics Combined Purification Strategy

A Quantitative Glycomics and Proteomics Combined Purification Strategy

Published on: March 8, 2016

Quantitation by isobaric labeling: applications to glycomics.

James A Atwood¹, Lei Cheng, Gerardo Alvarez-Manilla

¹Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, Athens, Georgia 30602-4712, USA.

Journal of Proteome Research

|December 1, 2007

Summary

Quantitation by isobaric labeling (QUIBL) enables precise measurement of glycan changes. This novel method facilitates comparative glycomics, offering insights into cell development and disease by quantifying glycan expression differences.

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Area of Science:

Glycomics
Mass Spectrometry
Biochemistry

Background:

Glycosylation patterns (glycomics) are crucial for understanding cell development and pathology.
Quantifying glycan expression changes between biological samples is a significant challenge in glycomics.

Purpose of the Study:

To introduce a novel strategy, quantitation by isobaric labeling (QUIBL), for facilitating comparative glycomics.
To enable accurate quantification of glycan expression differences in biological samples.

Main Methods:

QUIBL utilizes permethylation with isotopically labeled reagents (e.g., (13)CH 3I or (12)CH 2DI) to create isobaric glycan derivatives.
Separation and quantitation are achieved by exploiting the precise mass differences (0.002922 Da per methylation site) at high mass resolution (approx. 30000 m/Delta m).

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Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

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A Quantitative Glycomics and Proteomics Combined Purification Strategy

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Published on: March 8, 2016

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

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Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

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Main Results:

QUIBL demonstrated relative quantitation of N-linked oligosaccharides from a standard glycoprotein and human serum over a 2-order of magnitude linear dynamic range.
The method successfully allowed for the relative quantitation of isomeric glycans.
Applied to murine embryonic stem cell differentiation, QUIBL quantified glycomic changes during the transition to embryoid bodies.

Conclusions:

QUIBL is a powerful new technique for comparative glycomics, enabling precise quantification of glycan expression.
This method has broad applications in studying biological processes, disease mechanisms, and cell differentiation.