Protein purification can involve challenging separations of complex mixtures that may not resolve target proteins in a single chromatographic step. Such cases may require the use of multidimensional chromatography. Multidimensional chromatography combines a series of columns, each employing different separation mechanisms, into a single separation. Most common protein purification techniques can be incorporated into a multidimensional chromatography scheme, and such schemes provide great benefit for proteins purified on a routine basis. Typically, a multidimensional chromatographic scheme consists of an affinity purification step based on binding to a protein tag, such as multiple histidine (His) or glutathione-S-transferase (GST), followed by one or more additional resolution steps using ion exchange, size exclusion, or hydroxyapatite columns. To streamline the combination of such disparate chromatographic methods, an automated chromatography system with a flexible design and versatile control software is essential.
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