Preparation of a monolithic column for weak cation exchange chromatography and its application in the separation of biopolymers

作者：Wei YM^*; Huang XD; Liu RX; Shen YH; Geng XD

来源：Journal of Separation Science, 2006, 29(1): 5-13.

DOI：10.1002/jssc.200500210

• bioseparation
• ion-exchange chromatography
• liquid chromatography
• monolithic column
• renaturation of recombinant human interferon gamma

A procedure for the preparation of a monolithic column for weak cation exchange chromatography was presented. The structure of the monolithic column was evaluated by mercury intrusion. The hydrodynamic and chromatographic properties of the monolithic column - such as back pressures at different flow rates, effects of pH on protein retention, dynamic loading capacity, recovery, and stability - were determined under conditions typical for ion-exchange chromatography. The prepared monolithic column might be used in a relatively broad pH range from 4.0 to 12.0 and exhibited an excellent separation to five proteins at the flow rates of both 1.0 and 8.0 mL/min, respectively. In addition, the prepared column was first used in the purification and simultaneous renaturation of recombinant human interferon gamma (rhIFN-gamma) in the extract solution with 7.0 mol/L guanidine hydrochloride. The purity and specific bioactivity of the purified rhIFN-gamma in only one chromatographic step were obtained to be 93% and 7.8 x 10(7) IU/mg, respectively.

• 出版日期2006-1
• 单位西北大学