Beaded agarose affinity chromatography of bovine fibroblast interferon.
Bovine fibroblast interferon (BoF-IFN), produced in bovine embryonic kidney cell cultures by priming and infection with bluetongue virus, was partially purified by controlled pore glass chromatography. The partially purified BoF-IFN then was subjected to beaded agarose affinity chromatography in 2 distinct fractions--1 after the addition of 1M NaCl and the other one after the addition of 1.5M NaCl containing 50% ethylene glycol. Analysis of fractions by sodium dodecyl sulfate/polyacrylamide-gel electrophoresis revealed a broad molecular weight range (14,900 to 27,900) for IFN eluted by 1M NaCl, and 2 discrete molecular weight ranges (16,000 to 19,500 and 28,300 to 34,000) for IFN eluted by 1.5M NaCl containing 50% ethylene glycol. The specific activity of the IFN eluted with 1.5M NaCl containing ethylene glycol was 2.85 X 10(6) U/mg of protein, compared with 5.7 X 10(5) U/mg of protein in the controlled pore glass-purified IFN.
Allen
G K
GK
Department of Veterinary Microbiology, College of Veterinary Medicine, University of Missouri, Columbia 65211.
Rosenquist
B D
BD
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D007372:Interferons / Q000032:analysis / Q000302:isolation & purification*; D008863:Microspheres; D008970:Molecular Weight
D007372:Interferons