A Simple and Efficient Method for the Purification of Human Gamma Interferon

Abstract

A preparative, sequential chromatographic procedure has been developed for the purification of human gamma interferon (HuIFN-γ). The four steps in the procedure are Controlled Pore Glass-adsorption chromatography, Concanavalin-A affinity chromatography, Heparin-Sepharose affinity chromatography and gel-filtration.

By virtue of the development of a coordinated effluent-affluent buffer scheme, eluants can also serve as loading buffers for the succeeding column. Consequently, crude HuIFN-γ preparations can be purified rapidly (approximately one week), easily, and is amenable to a semi-automated process.

The procedure has also been shown to be efficient. Here, as an example, it is reported that an overall purification of greater than 75,000-fold can be achieved, yielding a specific activity of 5.2·107 units/mg, and a recovery of 95.5%. In addition, the peak fraction, representing 37.8% of the applied activity, had a specific activity of l.0·108 units/mg protein and represents a purification of more than 145,000-fold.

An SDS-PAGE analysis of one such fraction indicated that approximately 40% of the final material was HuIFN-γ.