Current LC methods for the separation and analysis of peptides and proteins

Proteins are biological macromolecules essential for maintaining the structural and functional integrity of the cell. Protein malfunction is often responsible for the onset of severe disorders. For the investigation of individual proteins or protein complexes and the identification of interactions with other biological macromolecules, purified proteins are required. They can be obtained from a tissue or by their overexpression in bacteria, yeast, or cultured mammalian cells and purification consists in their isolation from the source based on differences in their chemico-physical properties. The final objective is to maintain the highest level of functionality with the fewest contaminants. From among a variety of purification strategies, column chromatography is the primary method used. This chapter reviews the chromatographic procedures most applied in protein purification and discusses the purification schemes, their advantages, disadvantages, and potential problems. It also reports a survey on a good number of applications.