Non-mitogenic form of recombinant Human acidic fibroblast growth factor (nrhaFGF) retained the cardio and neuro-protective characters of the wild-type aFGF while decreased the potential side effects like mitogenesis activity. In this paper, nrhaFGF was successfully expressed in Escherichia coli and yielded about 5% of the total soluble protein in the 10L fermentation tank. the succeedent chromatography, cation exchange and heparin affinity gained the purity 55.0% and 95.6% specifically, after desalting with Sephadex G 25, aggregation was found increased 2-4% after the lyophilization, and Sephadex G75 completely eliminated the aggregates while ultrafiltration with cellulose triacetate membrane left 2% aggregates in the samples. The chromatography method and the the aggregation removal method with size exclusion chromatography were easily scaled up and spade the work for the later clinical trials.