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Updated: Jul 7, 2026

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Published on: July 31, 2011
Peptides OFFGEL electrophoresis: a suitable pre-analytical step for complex eukaryotic samples fractionation
Jérôme Chenau1, Sylvie Michelland, Jonathan Sidibe
1Centre de Recherche INSERM/UJF U, Cibles Diagnostiques ou Thérapeutiques et Vectorisation des Drogues dans le Cancer du Poumon Institut Albert Bonniot, La Tronche, France. jerome.chenau@gmail.com
OFFGEL fractionation effectively separates complex proteomes like human plasma and secretome. This method is compatible with iTRAQ labeling, enhancing quantitative proteomics workflows.
Area of Science:
- Proteomics
- Biochemistry
- Analytical Chemistry
Background:
- Mammalian proteomes are highly complex, featuring proteins with a wide dynamic range.
- Effective analysis requires overcoming complexity through multiple fractionation steps.
- OFFGEL fractionation offers efficient and repeatable pre-fractionation of peptides.
Purpose of the Study:
- To evaluate OFFGEL fractionator technology for complex proteome separation.
- To assess compatibility with quantitative proteomic labeling techniques.
Main Methods:
- Utilized a 24-well OFFGEL device (pH 3-10) for peptide separation.
- Combined OFFGEL fractionation with reverse-phase liquid chromatography and MALDI TOF-TOF identification.
- Analyzed peptide repartition profiles using an algorithmic model based on amino acid charge.
Main Results:
- Successfully separated peptides from human secretome and plasma proteomes.
- Demonstrated the compatibility of OFFGEL technology with iTRAQ labeling.
- Explained peptide distribution based on charged amino acid content.
Conclusions:
- OFFGEL fractionation is a valuable tool for complex eukaryotic proteomes.
- The system is compatible with iTRAQ labeling for quantitative proteomic studies.
- OFFGEL fractionation enhances strategies for quantitative proteomics.
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