title: Immobilised metal-ion affinity chromatography purification of histidine-tagged recombinant proteins: a wash step with a low concentration of EDTA.
abstract: Immobilised metal-ion affinity chromatography (IMAC) is widely used for the purification of recombinant proteins in which a poly-histidine tag is introduced. However, other proteins may also bind to IMAC columns. We describe the use of a washing buffer with a low concentration of EDTA (0.5 mM) for the removal of proteins without histidine tag from IMAC columns. Four histidine-tagged recombinant proteins/protein complexes were purified to homogeneity from cell culture medium of insect cells by using an EDTA washing buffer. The presence of a low concentration of EDTA in washing buffers during IMAC may have a general application in the purification of histidine-tagged proteins.
authors.affiliation: Department of Medical Microbiology, University of Groningen, The Netherlands.
authors.forename: D F
authors.initials: DF
authors.lastname: Westra
authors.forename: G W
authors.initials: GW
authors.lastname: Welling
authors.forename: D G
authors.initials: DG
authors.lastname: Koedijk
authors.forename: A J
authors.initials: AJ
authors.lastname: Scheffer
authors.forename: T H
authors.initials: TH
authors.lastname: The
authors.forename: S
authors.initials: S
authors.lastname: Welling-Wester
mesh_terms: D002614:Chelating Agents / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D004492:Edetic Acid / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006639:Histidine / Q000737:chemistry*; D008670:Metals; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D002614:Chelating Agents; D008670:Metals; D011994:Recombinant Proteins; D006639:Histidine; D004492:Edetic Acid