ABSTRACT

Afnity chromatography is a method in which specic and reversible interactions are used for the isolation, separation, and purication of biomolecules from crude samples (Wilchek 2004). In afnity chromatography, a ligand molecule having specic recognition ability is bound on a matrix. The molecule to be puried is selectively captured by the ligand bound on the solid matrix by simply passing the solution containing the target through the chromatographic column under favorable conditions. The target molecules are then eluted by using proper elutants under conditions favoring elution, by adjusting the pH, ionic strength, or temperature, using specic solvents or competitive free ligands, so that the interactions between the ligand and the target are broken and the target molecules are obtained in a puried form. The afnity-based approach was introduced in 1968 by Cuatrecasas et al. (1968) to purify proteins, and today, it still represents one of the most powerful techniques available for purication of bioactive materials (Matejtschuk 1997). The general afnity-based approach

2.1 Introduction .................................................................................................... 39 2.2 Chromatography of Antibodies Using Macroporous Cryogels ......................40