Mouse Gamma Interferon Produced by a Cloned T-Cell Lymphoma. I. Purification and Physicochemical Characterization

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Abstract
Mouse gamma interferon (IFN-γ) was produced from a cloned T-cell lymphoma, L12-R4, stimulated with phorbol myristic acetate (PMA) at a concentration of 2 × 10-7 M/ml. Absorption of crude IFN-γ on silicic acid resulted in a 100-fold concentration and 10-fold purification. After application of this IFN-γ 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 α-methyl-mannoside. The specific activity of the recovered IFN-γ was about 3.6 × 106 mg/ml of protein with 11,390-fold purification. Further investigations with Phenyl-Sepharose CL-4B and Chromatofocusing revealed that L12-R4 IFN-γ is highly hydrophobic and has a pi around 5.4. Thus, by different types of chromatography procedures, L12-R4 IFN-γ was found to be highly homogeneous with physicochemical properties similar to those produced by normal T-lymphocytes.