title: High-level production of murine interleukin-5 (IL-5) utilizing recombinant baculovirus expression. Purification of the rIL-5 and its use in assessing the biologic role of IL-5 glycosylation.
abstract: We describe here a recombinant baculovirus expression system useful for high level production of murine recombinant interleukin-5 (rIL-5). In addition, we describe a single-step technique of purification of the rIL-5 from the baculovirus-infected Sf9 cell supernatants, using an anti-IL-5 affinity column. The baculovirus-derived rIL-5 has physical properties and functional activities in various lymphoid cell assays similar to those of natural T cell-derived IL-5 and reacts with anti-IL-5 antibodies. Finally, the rIL-5 is similar to natural T cell-derived IL-5 in manifesting heterogeneous glycosylation; however, glycosylation does not appear to be necessary for biologic function, at least in a lymphoid cell proliferation assay.
authors.affiliation: Department of Medicine, University of Alberta, Edmonton, Canada.
authors.forename: D Y
authors.initials: DY
authors.lastname: Kunimoto
authors.forename: K C
authors.initials: KC
authors.lastname: Allison
authors.forename: C
authors.initials: C
authors.lastname: Watson
authors.forename: T
authors.initials: T
authors.lastname: Fuerst
authors.forename: G D
authors.initials: GD
authors.lastname: Armstrong
authors.forename: W
authors.initials: W
authors.lastname: Paul
authors.forename: W
authors.initials: W
authors.lastname: Strober
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002846:Chromatography, Affinity; D006031:Glycosylation; D007070:Immunoglobulin A / Q000096:biosynthesis; D007313:Insecta; D015848:Interleukin-5 / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D015183:Restriction Mapping; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014162:Transfection
chemical_list: D007070:Immunoglobulin A; D015848:Interleukin-5; D011994:Recombinant Proteins