Two-dimensional electrophoresis in proteome expression analysis

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-04, Vol.849 (1), p.190-202
• Main Author: LOPEZ, J. L
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Electrophoresis, Gel, Two-Dimensional - methods; Expression proteomics; Fundamental and applied biological sciences. Psychology; General pharmacology; Medical sciences; Pharmacology. Drug treatments; Proteome - analysis; Proteome - chemistry; Proteomics - methods; Reproducibility of Results; Sample preparation; Two-dimensional electrophoresis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2006.11.049

Cell proteomes are complex, given they consist of several thousand proteins. Two-dimensional electrophoresis (2DE) is unique not only for its ability to simultaneously separate thousands of proteins but also for detecting post- and co-translational modifications, which cannot be predicted from genome sequences. This review will describe the protocols applied to prepare 2D gels properly, and analyse and summarise the major challenges for successful proteome analysis using 2DE, i.e. the ability to analyse very alkaline, hydrophobic and/or low or high M r proteins with high resolution and the ability to detect minor components. Challenges involving sample preparation and solubilisation prior to the first dimension IEF/IPG step will be studied in depth. Sample preparation is crucial in 2DE studies and greatly influences other stages of the technique. It is the aim of this review not only to describe the challenges and limitations of 2DE but also to suggest the avenues, the evolution, the potential and the future of 2DE in proteomics.