title: Expression, renaturation and purification of recombinant human interleukin 4 from Escherichia coli.
abstract: The lymphokine human interleukin 4 (IL-4) has been expressed from a plasmid in the cytoplasm of Escherichia coli. Advantage has been taken of insolubility of the human IL-4 in E. coli for rapid purification of this protein in only a few steps. We describe extraction and renaturation procedures which solubilize human IL-4 yielding biologically active protein. The protein was purified to homogeneity by one passage over a gel-filtration column. The refolded human IL-4 was characterized by N-terminal sequence analysis, amino acid analysis and bioassays. The refolded E. coli-derived human IL-4 has biological activity on T and B cells and binds to the human IL-4 receptor, comparable to mammalian expressed human IL-4, indicating that the protein is folded correctly.
authors.affiliation: DNAX Research Institute, Palo Alto, California 94304.
authors.forename: A
authors.initials: A
authors.lastname: van Kimmenade
authors.forename: M W
authors.initials: MW
authors.lastname: Bond
authors.forename: J H
authors.initials: JH
authors.lastname: Schumacher
authors.forename: C
authors.initials: C
authors.lastname: Laquoi
authors.forename: R A
authors.initials: RA
authors.lastname: Kastelein
mesh_terms: D000596:Amino Acids / Q000302:isolation & purification; D001667:Binding, Competitive; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D015847:Interleukin-4; D007378:Interleukins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D046911:Macromolecular Substances; D010957:Plasmids; D011489:Protein Denaturation; D011971:Receptors, Immunologic / Q000187:drug effects; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D012995:Solubility; D014170:Transformation, Genetic
chemical_list: D000596:Amino Acids; D007378:Interleukins; D046911:Macromolecular Substances; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D015847:Interleukin-4