title: The construction and characterization of a biologically active recombinant IL-2 containing a lysine-rich C-terminal extension.
abstract: A polylysine extended gibbon IL-2 (IL-2-L) was constructed by the addition of a lysine-rich oligopeptide, Gly3-(Lys-Lys-Asp)3-Leu-Glu to the C terminus of gibbon IL-2 by using rDNA technology. The bioactivity of the hybrid molecule was comparable to that of normal human rIL-2 in assays measuring T cell proliferation, and in generation of lymphokine-activated killer cells from PBL. Furthermore, the addition of the lysine-rich segment made the molecule more amenable to the chemical derivatization of amino groups. After biotinylation, IL-2-L retained a greater proportion of its bioactivity than normal rIL-2. In addition, biotinylated IL-2-L was capable of simultaneously binding to cell surface IL-2R and streptavidin. Thus, the addition of the lysine-rich oligopeptide facilitated the generation of an active form of biotinylated IL-2 which acts as a bridge between IL-2R-positive cells and avidin-coupled reagents and affinity supports. Such a selective means of labeling and binding IL-2-responsive cells may have great practical utility for IL-2-based immunotherapy.
authors.affiliation: E. I. Du Pont and Co., Glenolden Laboratory, PA 19036.
authors.forename: Y
authors.initials: Y
authors.lastname: Lin
authors.forename: R J
authors.initials: RJ
authors.lastname: Robb
authors.forename: J E
authors.initials: JE
authors.lastname: Gray
authors.forename: P
authors.initials: P
authors.lastname: Simon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D001667:Binding, Competitive; D001710:Biotin / Q000378:metabolism; D002460:Cell Line; D006801:Humans; D006922:Hylobates; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D011107:Polylysine; D015375:Receptors, Interleukin-2 / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology; D019809:Streptavidin
chemical_list: D001426:Bacterial Proteins; D007376:Interleukin-2; D015345:Oligonucleotide Probes; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D011107:Polylysine; D001710:Biotin; D019809:Streptavidin