Role of chromatographic techniques in proteomic analysis

Proteomics, the characterization of the proteome, is conceptually simple but technically challenging. Development of such technologies as mass spectrometry, multidimensional protein separation, and DNA sequencing has allowed the new field of proteomics to flourish. Proteomic analysis relies on a set of techniques chosen on the basis of the biological question. In any proteomic analysis, the first and most important task is the separation of a complex protein mixture, i.e. the proteome. Chromatography, one of the most powerful methods of separation, employs one or more inherent characteristics of a protein—its mass, isoelectric point, hydrophobicity or biospecificity. This review emphasizes high-performance liquid chromatography as an integrated part of technologies used to study the proteome, discusses the capabilities and limitations of current instruments, and highlights the potential of multidimensional liquid chromatography in proteomic analysis.