Dye-Ligand Affinity Chromatography for Protein Separation and Purification

Affinity chromatography has proven to be the most effective technique for the purification and separation of proteins from complex mixtures (1 ). Although affinity adsorbents based on biological ligands such as immobilized antibodies, lectins and nucleotide cofactors appear to be highly successful, their use at a preparative scale is limited because of their instability, expense, and low capacity (1 ). Synthetic affinity ligands, such as reactive chlorotriazine dyes, have become an integral part of affinity-based protein purification methods for a number of reasons. The dyes are inexpensive, chemical immobilization of the dyes to the matrix is easy and the resultant dye-adsorbents are resistant to chemical or biological degradation, the protein binding capacity is high and far exceeds the binding capacity exhibited by biological ligands (1 ,2 ). The main disadvantage of reactive chlorotriazine dyes appears to be their moderate selectivity, which may limit their use. On the other hand, their lack of selectivity, in certain circumstances, may be beneficial, as it circumvents the requirement for a different adsorbent for each putative purification (3 ).