Studies on purification of human gamma interferon: chromatographic behavior of accompanying IL2 and B-cell helper activity.
Gamma interferon (gamma IFN) was produced in human lymphocyte cultures stimulated by PHA. Titers were in the range of 10,000-30,000 U/ml. Crude gamma IFN was adsorbed on silicic acid, from which the antiviral activity was eluted by a buffer containing a high salt concentration and ethylene glycol. This treatment allowed quantitative recovery of gamma IFN with a specific activity of 5 X 10(5)-1 X 10(6) U/mg of proteins. IL2 and B-cell helper activities were adsorbed and eluted from silicic acid together with the antiviral activity. This finding might be of practical interest for the purification of these lymphokines, particularly IL2. Gamma interferon was further purified on Blue Sepharose to a specific activity of 2 X 10(7) U/mg. The resulting preparations still contained IL2 and B cell helper activities. However, taking advantage of the differences in apparent hydrophobicity and in isoelectric point, we were able to dissociate antiviral activity from lymphokines. Such dissociation should facilitate the study of the biological properties of human natural gamma IFN.
D000327:Adsorption; D001402:B-Lymphocytes / Q000276:immunology*; D006133:Growth Substances / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007371:Interferon-gamma / Q000302:isolation & purification*; D007376:Interleukin-2 / Q000302:isolation & purification*; D015847:Interleukin-4; D007525:Isoelectric Focusing; D008222:Lymphokines / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives; D012824:Silicic Acid
D006133:Growth Substances; D007376:Interleukin-2; D008222:Lymphokines; D012824:Silicic Acid; D015847:Interleukin-4; C029483:Blue-Sepharose; D007371:Interferon-gamma; D012685:Sepharose