Dye affinity chromatography is a purification technique offering unique selectivities and high purification factors. Dye ligands may act as substrate analogs, offering affinity interactions with their corresponding enzymes. This chapter describes a dye ligand chromatography technique for purifying proteins from overexpression, in mammalian cell culture. The method begins with batch binding in order to rapidly select binding and elution conditions. Subsequently, gradient elution is employed to maximize the selectivity of the final packed bed chromatography method. Conditions for purification of a protein from mammalian cell culture on Cibacron blue are given with an accompanying sample chromatogram.
