High-level expression and purification of a nonmitogenic form of human acidic fibroblast growth factor in Escherichia coli.

To decrease the potential side effects of acidic fibroblast growth factor (aFGF) caused by its broad-spectrum mitogenic activity, a nonmitogenic form of aFGF (nhaFGF), which retained the cardio- and neuroprotective characters of the wild-type aFGF, was overexpressed in Escherichia coli. The expression level of nhaFGF was up to 25% of the total cellular protein. The expressed nhaFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. The mitogenic activity of the purified nhaFGF was decreased dramatically comparable to that of the wild-type aFGF (haFGF) detected by methylthiazoletetrazolium method. The purified recombinant nhaFGF was sufficiently prepared and sufficient for the following pharmacological study.