title: Purification and characterization of multiple components of human lymphoblastoid interferon-alpha.
abstract: Twenty-two components of human interferon-alpha (IFN-alpha) derived from Sendai virus-induced Namalwa cells were purified by sequential immunoadsorbent affinity chromatography using four monoclonal antibody affinity columns followed by ultrafiltration and reversed-phase high-performance liquid chromatography. The specific activity ranged from 0.2 to 2.6 x 10(8) IU/mg protein on Madin-Darby bovine kidney cells, 0.3 to 4.6 x 10(8) IU/mg protein on human WISH cells, and 10(4) to 7 x 10(5) units/mg protein on mouse L929 cells. The apparent molecular weights of the components ranged from 17,500 to 23,300 using nonreducing sodium dodecyl polyacrylamide-gel electrophoresis and 17,500 to 27,600 using reducing sodium dodecyl polyacrylamide-gel electrophoresis. The amino-terminal amino acid sequences were similar among the components as well as to those reported for the cloned human IFN-alpha genes (Pestka, S. (1986) Methods Enzymol. 119, 3-14). However, four components, f, i, l, and m, have amino-terminal amino acid sequences which appear to be unique when compared to those predicted from the cDNA clones. One component, pre-a, has a potential N-linked glycosylation site on the Asn of residues 2 through 4, Asn-Leu-Ser.
authors.affiliation: Division of Cytokine Biology, Food and Drug Administration, Bethesda, Maryland 20892.
authors.forename: K C
authors.initials: KC
authors.lastname: Zoon
authors.forename: D
authors.initials: D
authors.lastname: Miller
authors.forename: J
authors.initials: J
authors.lastname: Bekisz
authors.forename: D
authors.initials: D
authors.lastname: zur Nedden
authors.forename: J C
authors.initials: JC
authors.lastname: Enterline
authors.forename: N Y
authors.initials: NY
authors.lastname: Nguyen
authors.forename: R Q
authors.initials: RQ
authors.lastname: Hu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013045:Species Specificity; D014462:Ultrafiltration
chemical_list: D000911:Antibodies, Monoclonal; D016898:Interferon-alpha