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A Quantitative Glycomics and Proteomics Combined Purification Strategy
Published on: March 8, 2016
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Quantitative analysis of glycated proteins
Feliciano Priego-Capote1, María Ramírez-Boo, Francesco Finamore
1Translational Biomarker Group (TBG), Department of Human Protein Sciences, University Medical Centre, University of Geneva , 1211 Geneva 4, Switzerland.
Journal of Proteome Research
|January 15, 2014
Summary
This study introduces glycation isotopic labeling (GIL), a new method for analyzing glycated proteins (GP) in biological samples. GIL uses isotopic glucose labeling for accurate quantification and qualitative analysis of GPs, reflecting glycemic states.
Area of Science:
- Biochemistry
- Proteomics
- Analytical Chemistry
Background:
- Glycated proteins (GP) are important biomarkers for metabolic diseases.
- Accurate quantification of GPs in complex biological samples remains challenging.
Purpose of the Study:
- To develop a robust method for large-scale qualitative and quantitative analysis of glycated proteins.
- To enable reliable assessment of glycated protein levels in biological fluids and cell lysates.
Main Methods:
- Glycation isotopic labeling (GIL) using differential labeling with [(13)C6]-glucose.
- Enzymatic digestion with endoproteinase Glu-C.
- Mass spectrometry (MS) precursor scanning to detect doublet signals of glycated peptides.
- High-energy collisional dissociation (HCD-MS2) and collision-induced dissociation (CID-MS3 neutral loss scan) for qualitative analysis.
Main Results:
- The GIL method allows for the detection of doublet signals for glycated peptides with a mass shift dependent on charge state and glycation sites.
- Intensity ratios of doublet signals provide quantitative information on glycated proteins.
- The method is applicable to complex biological samples like plasma and red blood cells.
Conclusions:
- The glycation isotopic labeling (GIL) protocol offers a comprehensive approach for analyzing glycated proteins.
- This method facilitates the quantitative assessment of glycated proteins, correlating with glycemic states.
- GIL is a valuable tool for large-scale proteomic studies involving glycated proteins.
