Partial purification and characterization of feline gamma-like interferon.
Feline interferon (IFN) was produced from spleen cells stimulated by Staphylococcus enterotoxin A (SEA). The IFN was purified by a three-step procedure using controlled pore glass adsorption chromatography, concanavalin A (ConA)-agarose column chromatography and gel filtration. The final product of these procedures which had activity of an IFN appeared as a single peak of activity with molecular weight of approximately 50,000. It was sensitive to acid and heat, suggesting that the isolated material was a gamma IFN. The total recovery of feline gamma IFN was 8.2%. Specific activity was 2.95 X 10(4) unit/micrograms protein and was concentrated 2.8 X 10(4) times. This preparation of purified feline gamma IFN was destroyed completely by 0.1% sodium dodecyl sulfate within 20 min. As an inducer of feline gamma IFN, SEA appeared to produce a more uniform IFN product than did ConA. Further, the presence of 10% ethylene glycol in the sample applied to ConA-agarose column as well as its absence in the elution buffer was effective in reducing contaminating acid- and heat-resistant IFNs in the preparation.
D000818:Animals; D002415:Cats; D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D003208:Concanavalin A / Q000494:pharmacology; D004768:Enterotoxins / Q000494:pharmacology; D007371:Interferon-gamma / Q000096:biosynthesis / Q000302:isolation & purification*; D013154:Spleen / Q000276:immunology; D010948:Viral Plaque Assay
D004768:Enterotoxins; D003208:Concanavalin A; C026066:enterotoxin A, Staphylococcal; D007371:Interferon-gamma