Open AccessPurification and characterization of human interleukin‐1β expressed in recombinant Escherichia coli
Author(s) -
WINGFIELD Paul,
PAYTON Mark,
TAVERNIER Jean,
BARNES Marjory,
SHAW Alan,
ROSE Keith,
SIMONA Marco G.,
DEMCZUK Stephen,
WILLIAMSON Karen,
DAYER JeanMichel
Publication year - 1986
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1986.tb10066.x
Subject(s) - escherichia coli , recombinant dna , microbiology and biotechnology , in vitro , peripheral blood mononuclear cell , biochemistry , biology , peptide sequence , amino acid , chemistry , lymphocyte , gene , immunology
The high‐level expression of human interleukin‐1β in Escherichia coli is described. The protein contributes about 12% of the total cell protein and is associated with the soluble cytoplasmic fraction of the cell. A method for the purification of the protein is given which is based on anion‐ and cation‐exchange chromatographies. The isolated protein, shown to be homogeneous by several analytical methods, has been characterized by amino acid analysis, N‐ and C‐terminal sequence analysis and analytical centrifugation. The protein is biologically active as demonstrated by two different in vitro assays, namely, the mononuclear cell factor (IL‐1/MCF) assay and lymphocyte activating factor (IL‐1/LAF) assay. The specific activities determined with the IL‐1/MCF and IL‐1/LAF assays, are 2 × 10 7 and 4 × 10 7 units mg −1 , respectively.