Affinity Monolith Chromatography: Principles and Recent Developments


Michelle J. Yoo and David S. Hage


Affinity monolith chromatography (AMC) is a type of liquid chromatography (LC) that combines the use of a monolithic support with a biologically related binding agent as the stationary phase. In this chapter we describe the basic principles behind affinity chromatography and examine the types of supports and ligands that have been employed in this method. Several recent applications of AMC are discussed, including its use with bioaffinity chromatography, immunoaffinity chromatography, dye-ligand affinity chromatography, immobilised metal-ion affinity chromatography (IMAC) and the study of biological interactions. Closely related applications of monoliths in the areas of affinity-based chiral separations, enzyme reactors and microfluidic devices are also examined. Finally, the advantages and disadvantages of AMC versus other types of affinity chromatography or LC are considered.


1.1 Introduction


1.1.1 Basic Principles of Affinity Chromatography Affinity chromatography is a chromatographic technique that involves the use of a biologically related ligand as a stationary phase. This method uses reversible and selective interactions that characterise many biological systems, such as the binding of an antibody with an antigen or an enzyme with a substrate or inhibitor. Affinity chromatography makes use of the high selectivity found in many of these interactions by placing such binding agents within a column and using this column to isolate, measure or examine the binding of complementary compounds to this agent [1–3]. The binding agent in this case is called the ‘affinity ligand’ and can be immobilised onto the support through various methods, including adsorption, covalent immobilisa- tion or entrapment [1, 2]. In this chapter we examine the basic principles of affinity


Monolithic Chromatography and its Modern Applications, edited by Perry G. Wang. # 2010 ILM Publications, a trading division of International Labmate Limited.


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