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

✍ Scribed by Yinmao Wei; Xiaodong Huang; Ruixian Liu; Yehua Shen; Xindu Geng

Publisher
John Wiley and Sons
Year
2006
Tongue
English
Weight
516 KB
Volume
29
Category
Article
ISSN
1615-9306

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✦ Synopsis

Abstract

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.

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