title: Purification of recombinant histidine-tag streptolysin O using immobilized metal affinity expanded bed adsorption (IMA-EBA).
abstract: In this report, we describe the recombinant SLO expression as a fusion protein with a C-terminal hexahistidine tag and its purification using immobilized metal affinity expanded bed adsorption (STREAMLINE(trade mark) Chelating). In order to facilitate downstream processing of the purification, an efficient fermentation process was developed focusing on the achievement of high yields of soluble protein. The purification strategy resulted in a 40% recovery of active recombinant SLO and the protein was purified eight-fold. SDS-PAGE and Western-blot analysis of the purified protein revealed the presence of a 75 Mr form, which was the estimated relative Mass of the recombinant SLO.
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Unitat de Bioquímica de Medicina, Universitat Autònoma de Barcelona, Edifici M, 08193 Bellaterra, Barcelona, Spain. sandra.campubri@uab.es
authors.forename: Sandra
authors.initials: S
authors.lastname: Camprubí
authors.forename: Marc
authors.initials: M
authors.lastname: Bruguera
authors.forename: Francesca
authors.initials: F
authors.lastname: Canalias
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification; D015153:Blotting, Western; D002614:Chelating Agents / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005285:Fermentation; D006639:Histidine; D009532:Nickel / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D013301:Streptolysins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; D002614:Chelating Agents; D011993:Recombinant Fusion Proteins; D013301:Streptolysins; C025964:streptolysin O; D006639:Histidine; D009532:Nickel