65] Purification of human fibroblast interferon by adsorption to controlled-pore glass and zinc-chelate chromatography

Abstract

Publisher Summary This chapter presents purification techniques for human interferons yield in either small quantities of high purity, for example, sufficient for determination of specific activities, or larger quantities of low purity. However, methods are described for purification of larger quantities of completely pure interferon. The procedure described yields large quantities of pure human fibroblast interferon in two steps. The overall recovery is at least 60% of the starting materials, and the specific activity of the end product varies from 1.5 to 3.1 x 10 9 reference units/mg, averaging about 2 x 10 9 reference units per milligram of protein. The uniqueness of the method is that the two steps yield a pure product when used in combination with each other, but not when either step is used in combination with other purification procedures, such as concanavalin A-Sepharose or phenyl-Sepharose chromatography. The first step involves the partial purification of crude human fibroblast interferon by controlled-pore glass (CPG) adsorption. The second step consists of a modified and large-scale adaptation of the zinc-chelate chromatographic purification method as a first step in the purification of crude interferon.