title: Preparation, analysis and use of an affinity adsorbent for the purification of GST fusion protein.
abstract: Methods are presented for the preparation, ligand density analysis and use of an affinity adsorbent for the purification of a glutathione S-transferase (GST) fusion protein in packed and expanded bed chromatographic processes. The protein is composed of GST fused to a zinc finger transcription factor (ZnF). Glutathione, the affinity ligand for GST purification, is covalently immobilized to a solid-phase adsorbent (Streamline). The GST-ZnF fusion protein displays a dissociation constant of 0.6 x 10(-6) M to glutathione immobilized to Streamline. Ligand density optimization, fusion protein elution conditions (pH and glutathione concentration) and ligand orientation are briefly discussed.
authors.affiliation: Department of Chemical Engineering, Monash University, Clayton, Victoria, Australia.
authors.forename: Gareth M
authors.initials: GM
authors.lastname: Forde
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase