Principles of Chromatography

Chromatography in its many different forms is an indispensable separation technique for purification or analysis of one or multiple components from a mixture that may contain a variety of simple as well as complex molecules. This unit is focused on a specific type of chromatography known as liquid chromatography (LC), with particular reference to separation of proteins. The theory of chromatography is discussed in the context of partition of the components of a mixture between a solid stationary phase and a liquid mobile phase. Although very useful for chromatographic separation of small molecules, considerable deviation is observed in the case of separation of proteins. The principles of three commonly used chromatographic techniques for separation of proteins, such as size exclusion, ion exchange, and affinity chromatography, are discussed. The unit is expected to be useful for biologists and biochemists, as well as engineers involved in downstream bioprocessing. © 2016 by John Wiley & Sons, Inc. Keywords: chromatography; liquid chromatography; theory of chromatography; plate model; protein purification; size exclusion chromatography; gel filtration; ion exchange chromatography; affinity chromatography