Abstract
This chapter discusses the production and purification of mouse immune interferon. Crude murine immune interferon preparations contain a variety of lymphokines and biologically active molecules other than murine gamma-interferon (Mu-IFN-γ). The execution of in vivo experiments demands large quantities of Mu-IFN-γ, which should be largely devoid of these substances. The investigations on the biochemical properties of the Mu-IFN-γ molecules, and the generation of both polyclonal and monoclonal antibodies (e.g., in rat), require highly purified antigen preparations. Methods for obtaining such preparations are described in this chapter. The highest levels of Mu-IFN-γ production are obtained with the lysate of S. aureus 13N2909. On average, a level of 10 3.1 -10 4.3 units/ml is reached after 72 hr of incubation. This crude Mu-IFN-y is purified by a sequence of silicic acid adsorption/elution, affinity chromatography on Concanavalin A (Con A)-Sepharose and anion-exchange chromatography on fast protein liquid chromatography (FPLC) mono Q. The average purification factor, recovery, and specific activity obtained after these different steps are summarized in the chapter.
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