Use of ZetaPrep cartridge for the purification of human recombinant interleukin 1 beta.
Zetaprep mass ion-exchange media represent a rapid and efficient chromatographic tool in the separation of proteins, in place of the conventional agarose or cellulose-based gels. We adopted this method, combined with classical steps, to purify to homogeneity human recombinant interleukin 1 beta (IL-1 beta) produced from E. coli and from S. cerevisiae. An anion exchanger QAE-ZetaPrep was used to achieve a rapid partial purification of both proteins. The IL-1 beta purification was completed by gel permeation chromatography on Sephadex G-50. When the protein was produced from yeast, an intermediate chromatographic step on a hydroxylapatite column was also necessary. The isolated proteins proved to be homogeneous by electrophoresis and amino acid analysis. The biological activity of IL-1 beta produced by E. coli is comparable to that of the natural protein, while the protein produced by yeast showed very low specific activity.
Casagli
M C
MC
Sclavo Research Centre, Siena, Italy.
Borri
M G
MG
D'Ettorre
C
C
Galeotti
C L
CL
Di Liegro
C
C
Ghiara
P
P
Antoni
G
G
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D007375:Interleukin-1 / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D007375:Interleukin-1; D011994:Recombinant Proteins