title: Rapid purification of histidine-tagged glutathione S-transferase fusion protein by metal chelate POROS perfusion chromatography media.
abstract: The use of POROS MC media allowed high speed column preparation, sample loading, and elution. Under these conditions the proteins of interest can be recovered under mild conditions to maintain maximum biological activity. The speed of the process also assures minimum degradation or post-extraction modifications of the protein of interest from exposure to naturally occurring proteases and other enzymes. It is evident from the analysis by polyacrylamide gel electrophoresis of the purified fusion protein and starting material that a high degree of purification is possible in a single high-speed separation on POROS MC. The high flow rate capabilities of POROS media offer several other significant advantages over conventional media. A few of these include high speed and systematic method optimization, rapid column preparation and sample loading, and fast time-saving chromatographic separations.
authors.affiliation: Boehringer Mannheim GmbH, Penzberg, Germany.
authors.forename: S B
authors.initials: SB
authors.lastname: Schmidbauer
authors.forename: O K
authors.initials: OK
authors.lastname: Strobel
mesh_terms: D002412:Cations / Q000378:metabolism; D002614:Chelating Agents / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006639:Histidine / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015203:Reproducibility of Results; D013053:Spectrophotometry
chemical_list: D002412:Cations; D002614:Chelating Agents; D011993:Recombinant Fusion Proteins; D006639:Histidine; D005982:Glutathione Transferase