title: Partial purification and characterization of bovine fibroblast interferon.
abstract: Bovine fibroblast interferon (BoF-IFN), produced in primary bovine embryonic kidney cell cultures after priming and infection with bluetongue virus, was purified by controlled pore glass (CPG) chromatography to a specific activity of 10(6) U/mg of protein, with 40% recovery of the original activity. The crude IFN was concentrated more than sevenfold during purification. This proved to be a relatively simple, practical method of obtaining sufficient quantities of partially purified natural BoF-IFN for further studies. The CPG-purified BoF-IFN was further concentrated by sequential ultrafiltration and was analyzed by sodium dodecyl sulfate/polyacrylamide-gel electrophoresis (SDS-PAGE). Interferon, recovered from denaturing conditions either by dialysis against phosphate-buffered saline solution or by dilution in cell culture medium containing 10% fetal bovine serum, migrated as a single stainable protein with molecular weight of 21,000 on analytic SDS-PAGE gels. Recovered IFN activity from preparative SDS-PAGE totalled 8.7% of that applied. Attempts to further purify CPG-purified BoF-IFN by zinc chelate affinity chromatography were unsuccessful.
authors: Department of Veterinary Microbiology, College of Veterinary Medicine, University of Missouri, Columbia 65211.
authors: G K
authors: GK
authors: Allen
authors: G D
authors: GD
authors: Grothaus
authors: B D
authors: BD
authors: Rosenquist
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002845:Chromatography; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D007372:Interferons / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D008863:Microspheres; D014462:Ultrafiltration
chemical_list: D007372:Interferons