Perspectives of immobilized-metal affinity chromatography.
Immobilized Metal-Affinity Chromatography (IMAC) represents a relatively new separation technique that is primarily appropriate for the purification of proteins with natural surface-exposed histidine residues and for recombinant proteins with engineered histidine tags or histidine clusters. Because the method has gained broad popularity in recent years, the main recent developments in the field of new sorbents, techniques and possible applications are discussed in this article. Advantages of the method and new prospects are described as well as the problems and concerns that appear when the method is to be used for production of pharmaceutical-grade proteins.
Gaberc-Porekar
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National Institute of Chemistry, Hajdrihova 19, SI-1000 Ljubljana, Slovenia. vladka.gaberc.porekar@ki.si
Menart
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D001665:Binding Sites; D002614:Chelating Agents; D002846:Chromatography, Affinity / Q000379:methods*; D003902:Detergents; D006639:Histidine; D008670:Metals; D011485:Protein Binding; D017510:Protein Folding; D011506:Proteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D012997:Solvents
D002614:Chelating Agents; D003902:Detergents; D008670:Metals; D011506:Proteins; D011994:Recombinant Proteins; D012997:Solvents; D006639:Histidine