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Preparation of a monolithic column for weak cation exchange chromatography and its application in the separation of biopolymers
Author(s) -
Wei Yinmao,
Huang Xiaodong,
Liu Ruixian,
Shen Yehua,
Geng Xindu
Publication year - 2006
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500210
Subject(s) - monolithic hplc column , chromatography , chemistry , ion chromatography , ion exchange , column chromatography , chromatography column , volumetric flow rate , high performance liquid chromatography , ion , organic chemistry , physics , quantum mechanics
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‐γ) in the extract solution with 7.0 mol/L guanidine hydrochloride. The purity and specific bioactivity of the purified rhIFN‐γ in only one chromatographic step were obtained to be 93% and 7.8×10 7 IU/mg, respectively.