Title: A rapid single-step purification method for human interferon-gamma from isolated Escherichia coli inclusion bodies
Authors: Haelewyn, J ×
De Ley, Marc #
Issue Date: Jan-1995
Publisher: Academic press aust
Series Title: Biochemistry and molecular biology international vol:37 issue:6 pages:1163-1171
Abstract: A fast purification method for recombinant human interferon-gamma, produced in E. coli, was elaborated. Human IFN-gamma accumulated in the cytoplasm of E. coli cells as inclusion bodies (IB). After lysis, the IB were isolated from the cell debris by means of a density gradient ultracentrifugation, and solubilized in 6 M guanidine hydrochloride. The subsequent refolding step was optimized for a maximal recovery of the biologically active dimer. Refolded IFN-gamma was then purified to homogeneity in a single cation exchange chromatographic step, yielding 12.5 mg protein per liter E. coil culture. The dimeric nature of the refolded protein was visualized by means of interchain cross-linking. In a subsequent Western blot the resulting derivative was recognized by a panel of five monoclonal antibodies, indicating that those epitopes on the protein surface remained unaffected upon cross-linking.
ISSN: 1039-9712
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Biochemistry, Molecular and Structural Biology Section
× corresponding author
# (joint) last author

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