Liquid Chromatography - Affinity Chromatography

Affinity chromatography can be defined as a liquid chromatographic method in which a biological agent or biomimetic ligand is used for the selective retention of complementary compounds. This form of liquid chromatography was originally used by Starkenstein in 1910 for the purification of amylase through the use of starch as a solid support. This method continued to slowly develop over the next 50 years. However, it was not until the 1960s that suitable supports like beaded agarose, as developed by Hjerten, became available as well as relatively simple immobilization techniques for these supports. An important advancement in this latter area was a report in 1967 by Axen, Porath, and Ernback in which the cyanogen bromide method for protein and peptide immobilization was first reported. This approach was then used in 1968 by Cuatrecasas, Anfinsen, and Wilchek to purify enzymes through the use of immobilized enzyme inhibitors. It was also at this time that the term ‘affinity chromatography’ was proposed to describe this technique. Affinity chromatography is relatively simple to perform and is a powerful tool for the separation of biological macromolecules. The high selectively of this approach often allows single-step purification strategies to be developed, even when working with dilute and highly complex mixtures. This simplicity and the variety of ligands that can be used with this approach have made it an important tool in process-scale separations. However, modern affinity chromatography also plays an important role in the analysis and study of biological systems. For instance, most forms of chiral....