Two-step Purification of Recombinant Human Beta Nerve Growth Factor(?-rhNGF) Secreted by CHO Cells with Chromatographic Method
- VernacularTitle:两步柱色谱法纯化CHO表达的重组人?神经生长因子(?-rhNGF)
- Author:
Jing JIANG
;
Shu-Ping YU
;
Gui-Xiang JIANG
;
Chang-Hai WANG
;
- Publication Type:Journal Article
- Keywords:
Nerve growth factor(NGF) Purification Ion-exchange Reversed-phase chromatography
- From:
China Biotechnology
2006;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
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(? –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 ? –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 ? –NGF bought from Sigma,which indicated that there is no loss of biological activity through the isolation process.The conclusion suggested that an economical isolation method of recombinant human ? –NGF could be practiced on the industrial process of purification.
