title: Micropreparative purification of recombinant human interleukin-2.
abstract: Recombinant Interleukin-2 (IL-2) is expressed in E. coli as insoluble aggregates; a protocol has been developed for solubilization, renaturation and purification of IL-2 from such aggregates at the 5-10-mg level. IL-2 aggregates were isolated from soluble proteins by centrifugation, subjected to a 1 M guanidine hydrochloride wash and a butan-1-ol wash (the latter to remove lipid), dissolved in 8 M guanidine hydrochloride-10 mM dithiothreitol and partly purified by gel permeation chromatography. Refolding/oxidation was then performed by dilution into Tris-HCl, pH 8.5 containing 1.5 microM copper sulphate to accelerate autoxidation. Final purification was by successive cation-exchange and reversed-phase high-performance liquid chromatographic steps, yielding over 99.5% pure IL-2 with an overall recovery of 20%.
authors.forename: M P
authors.initials: MP
authors.lastname: Weir
authors.forename: J
authors.initials: J
authors.lastname: Sparks
authors.forename: A M
authors.initials: AM
authors.lastname: Chaplin
mesh_terms: D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007376:Interleukin-2 / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate
chemical_list: D007376:Interleukin-2; D011994:Recombinant Proteins; D012967:Sodium Dodecyl Sulfate