Preparation of Weak Anion Exchange Chromatographic Packings Based on Monodisperse Polymer Beads and their Application in the Separation of Bioploymers

Abstract

Based on the 8.0 µm monodisperse, macroporous poly‐(glycidylmethacrylate‐co‐ethylene‐dimethacrylate) beads (P_GMA/EDMA), a new hydrophilic weak anion‐exchange (WAX) stationary phase for HPLC was synthesized by a new chemically modified method. The stationary phase was evaluated in detail to determine its ion‐exchange properties, separability, reproducibility, hydrophilicity, and the effect of column loading and pH on the separation and retention of proteins. It was found to have an ion‐exchange chromatographic (IEC) retention mechanism. The dynamic protein loading capacity of the synthesized SAX packings for BSA was 27.4 mg · g⁻¹. Four proteins were separated within 6.0 minutes using the synthesized WAX resin. The WAX resin was also used for the rapid separation and purification of recombinant human granulocyte colony stimulating factor (rhG‐CFS) from the coarse extract solution with only one step. The purity of the purified rhG‐CFS was more than 95%.

Keywords:

• Monodisperse poly(glycidyl methacrylate‐co‐ethylenedimethacrylate) resins
• Weak anion exchange chromatography
• Protein separation
• Recombinant human granulocyte colony stimulating factor

Acknowledgments

Financial support of this work by the National Scientific Foundation in China (No. 20465001), the Minister of Science and Technology Foundation (No. 2004CCA05000), and Program for New Century Excellent Talents in Chinese University (NCET‐04‐0986) are gratefully acknowledged. We also thank Prof. Tiansheng Su (Institute of Chemistry, Chinese Academy of Sciences, Beijing) for his helpful comments and suggestions.