色谱

色谱

• 研究论文 • 上一篇    下一篇

活性蓝F3GA固载的无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯亲和色谱填料的制备与应用

卜春苗1,王有贤1,龚波林1,2,阎超2

  

  1. 1.Key Laboratory of Energy &Chemical Engineering, Ningxia University, Yinchuan 750021, China;
    2.Unimicro (Shanghai) Technologies Co., Ltd., Shanghai 201203, China
  • 收稿日期:2007-09-04 修回日期:2007-11-22 出版日期:2008-05-30 发布日期:1984-03-25
  • 通讯作者: 龚波林

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

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摘要：

以3.0 μm无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂为基质,与三嗪染料活性蓝F3GA(Cibacron Blue F3GA)反应,制备出
一种固定化染料聚合物高效亲和色谱填料。考察了应用该填料时流动相中的盐离子浓度、有机溶剂及流速等对牛血清白蛋白(BSA)和
溶菌酶(Lys)保留行为的影响。实验结果表明,该染料亲和填料具有良好的色谱性能。利用前沿色谱法测定了染料与溶菌酶之间的表观
解离常数为5.26×10-5 mol/L。使用该填料成功地从鸡蛋清和小牛血清中分别分离纯化了Lys和BSA,十二烷基硫酸钠-聚丙烯酰胺凝胶
电泳(SDS-PAGE)分析显示Lys和BSA的纯度分别为95%和92%。

关键词: 纯度 , 蛋白质, 固定相, 染料亲和色谱, 无孔单分散亲水性树脂

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