Extraction of rIL‐2 inclusion bodies from Escherichia coli using cross‐flow filtration

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Abstract
A cross-flow membrane filtration process was developed for the recovery of rIL-2 inclusion bodies from homogenized Escherichia coli. The membrane extraction process was comprised of a two-step diafiltration followed by an extraction with 7 M GuHCl and a 40–fold dilution of the solubilized inclusion bodies into 0.01 M Tris–HCl, 0.035 M NaCl, pH 7.9. The first diafiltration was with a 0.03 M Tris–HCl, 5 mM ethylenediaminetetraacetic acid (EDTA), pH 8, followed by a diafiltration with 1.75 M GuHCl. All of the insoluble rIL-2 was retained behind the membrane, whereas a GuHCl wash solubilized approximately 15% of the rIL-2. The membrane process increased the yield of rIL-2 in the diluted extract by threefold as compared to a similar centrifuge process with a significant increase in purity as determined by reverse-phase high-performance liquid chromatography (HPLC). © 1994 John Wiley & Sons, Inc.