Chinese Journal of Chromatography

Chinese Journal of Chromatography

• Articles • Previous Articles     Next Articles

Preparation of affinity chromatographic packings by immobilization of Cibacron Blue F3GA based on monodispersed
non-porous hydrophilic poly (glycidylmethacry-co-ethylenedimethacrylate) particles and their application

BO Chunmiao1, WANG Youxian1, GONG Bolin1,2, YAN Chao2   

  1. 1.Key Laboratory of Energy &Chemical Engineering, Ningxia University, Yinchuan 750021, China;
    2.Unimicro (Shanghai) Technologies Co., Ltd., Shanghai 201203, China
  • Received:2007-09-04 Revised:2007-11-22 Online:2008-05-30 Published:1984-03-25

PDF

959

Abstract:

An affinity chromatographic packing was prepared by the immobilization of Cibacron Blue F3GA as dye
ligands on the base of monodispersed, 3.0 μm non-porous hydrophilic poly (glycidylmethacry-co-
ethylenedimethacrylate) particles. The effects of the ionic strength, organic solvent in the mobile phase and flow
rate on protein retention were investigated on the chromatographic stationary phase. The apparent dissociation
constant between the immobilized Cibacron Blue F3GA and lysozyme (Lys) was 5.26×10-5 mol/L which was evaluated by
frontal chromatography. Purifications of Lys from egg white and bovine serum albumin (BSA) from calf blood were
performed on the dye-ligand chromatographic column, separately, and the purities of the obtained fractions were
determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The purities of the purified Lys
and BSA were determined to be 95% and 92%, respectively.

Key words:
protein, dye-ligand affinity chromatography, purification
, stationary phase, monodispersed non-porous hydrophilic particles