Mouse gamma interferon produced by a cloned T-cell lymphoma. I. Purification and physicochemical characterization.
Mouse gamma interferon (IFN-gamma) was produced from a cloned T-cell lymphoma, L12-R4, stimulated with phorbol myristic acetate (PMA) at a concentration of 2 X 10(-7) M/ml. Absorption of crude IFN-gamma on silicic acid resulted in a 100-fold concentration and 10-fold purification. After application of this IFN-gamma to a Sephacryl S-200 column, 100% of the IFN activity was recovered in fractions containing molecular weights of 40 kD. When this material was applied to a Con A-Sepharose column, IFN activity was completely eluted with a linear gradient of alpha-methyl-mannoside. The specific activity of the recovered IFN-gamma was about 3.6 X 10(6) mg/ml of protein with 11,390-fold purification. Further investigations with Phenyl-Sepharose CL-4B and Chromatofocusing revealed that L12-R4 IFN-gamma is highly hydrophobic and has a pI around 5.4. Thus, by different types of chromatography procedures, L12-R4 IFN-gamma was found to be highly homogeneous with physicochemical properties similar to those produced by normal T-lymphocytes.
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000302:isolation & purification; D007525:Isoelectric Focusing; D008223:Lymphoma / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D013601:T-Lymphocytes; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate