Preparation of a monolithic column for weak cation exchange chromatography and its application in the separation of biopolymers.
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.
Institute of Modern Separation Science, Northwest University, Xi'an, China. ymwei@nwu.edu.cn
Yinmao
Y
Wei
Xiaodong
X
Huang
Ruixian
R
Liu
Yehua
Y
Shen
Xindu
X
Geng
D000327:Adsorption; D002412:Cations; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007371:Interferon-gamma / Q000737:chemistry / Q000302:isolation & purification*; D008768:Methylmethacrylates / Q000138:chemical synthesis / Q000737:chemistry*; D015394:Molecular Structure; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins
D002412:Cations; D008768:Methylmethacrylates; D011506:Proteins; D011994:Recombinant Proteins; C020347:GMA-EDMA; D007371:Interferon-gamma