Recent advances in peptide separation by multidimensional liquid chromatography for proteome analysis

Shotgun proteomics dominates the field of proteomics. The foundations of the strategy consist of multiple rounds of peptide separation where chromatography provides the bedrock. Initially, the scene was relatively simple with the majority of strategies based on some types of ion exchange and reversed phase chromatography. The thirst to achieve comprehensivity, when it comes to proteome coverage and the global characterization of post translational modifications, has led to the introduction of several new separations. In this review, we attempt to provide a historical perspective to separations in proteomics as well as indicate the principles of their operation and rationales for their implementation. Furthermore, we provide a guide on what are the possibilities for combining different separations in order to increase peak capacity and proteome coverage. We aim to show how separations enrich the world of proteomics and how further developments may impact the field.

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► A historical perspective to separations in proteomics is provided.
► Principles of operation and rationales for implementation will be discussed.
► A guide on what are the possibilities for combining separations will be provided.
► We aim to show how separations enrich the world of proteomics.