中国生物工程杂志

Nerve growth factor (NGF) was firstly discovered as a member of neurotrophin family, and the research and development of NGF has been lasting more than fifty years since its discovery. To this end, a two-step and high -yield chromatographic method which consists of cation ion-exchange chromatography and reversed-phase chromatography was reported to isolate recombinant human beta nerve growth factor (β -NGF) secreted by constructed Chinese hamster ovary cells (CHO/dhfr-) from the culture media. Through the process of purification, the purity of protein which was determined by SDS-PAGE and RP-HPLC has reached to 95%, and the recovery of β -NGF routed by RP-HPLC could be 70%. Furthermore, the biological activity of final purified protein evaluated by PC12 cells and dorsal root ganglia (DRG) exhibited the same performance as the standard protein of β -NGF bought from Sigma, which indicated that there is no loss of biological activity through the isolation process. This study's conclusion suggested that an economical isolation method of recombinant human β -NGF could be practiced on the industrial process of purification.