title: Protein purification by dye-ligand chromatography.
abstract: Dye-ligand chromatography has developed into an important method for large-scale purification of proteins. The utility of the reactive dyes as affinity ligands results from their unique chemistry, which confers both the ability to interact with a large number of proteins as well as easy immobilization on typical adsorbent matrices. Reactive dyes can bind proteins either by specific interactions at the protein's active site or by a range of non-specific interactions. Divalent metals participate in yet another type of protein-reactive dye interactions which involve the formation of a ternary complex. All of these types of interactions have been exploited in schemes for protein purification. Many factors contribute to the successful operation of a dye-ligand chromatography process. These include adsorbent properties, such as matrix type and ligand concentration, the buffer conditions employed in the adsorption and elution stages, and contacting parameters like flowrate and column geometry. Dye-ligand chromatography has been demonstrated to be suitable for large-scale protein purification due to their high selectivity, stability, and economy. Also, the issue of dye leakage and process validation of large-scale dye-ligand chromatography has been discussed. Reactive dyes have also been applied in high performance liquid affinity chromatographic techniques for protein purification, as well as non-chromatographic techniques including affinity partition, affinity membrane separations, affinity cross-flow filtration, and affinity precipitation.
authors.affiliation: Department of Chemical Engineering, Lehigh University, Bethlehem, Pennsylvania 18015.
authors.forename: P M
authors.initials: PM
authors.lastname: Boyer
authors.forename: J T
authors.initials: JT
authors.lastname: Hsu
mesh_terms: D000327:Adsorption; D002845:Chromatography / Q000379:methods*; D004396:Coloring Agents / Q000737:chemistry; D007700:Kinetics; D008024:Ligands; D011506:Proteins / Q000302:isolation & purification*
chemical_list: D004396:Coloring Agents; D008024:Ligands; D011506:Proteins