Purification and characterization of human interleukin‐1β expressed in recombinant Escherichia coli
Authors:
Open Access
- 1 November 1986
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 160 (3) , 491-497
Abstract
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 × 107 and 4 × 107 units mg−1, respectively.Keywords
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