OPTIMIZED PROCEDURES FOR PURIFICATION AND SOLUBILIZATION OF BASIC FIBROBLAST GROWTH-FACTOR INCLUSION-BODIES

The efficiency of purification of basic fibroblast growth factor (bFGF ) inclusion bodies using EDTA and nonionic detergents was improved fro m 25 to 40% by shifting the pH from 8.5 to strong alkaline conditions (pH 9.5 - 10.5). Complete dissolution of bFGF inclusion bodies by guan idinium hydrochloride (> 3 M) was independent of pH and the presence o f reducing agents. In contrast, solubilization of bFGF inclusion bodie s by urea was pH-dependent and increased in efficiency (e.g. from 0 to 100%) by increasing the pH (from pH 5.0 to 10.5 at 9 M urea). The pur ification and solubilization procedures are efficient for inclusion bo dy concentrations corresponding to 10 and 100 g per I dry cell weight, respectively.