Preparation, analysis and use of an affinity adsorbent for the purification of GST fusion protein.
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.
Forde
Gareth M
GM
Department of Chemical Engineering, Monash University, Clayton, Victoria, Australia.
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*
D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase