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 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
An affinity chromatographic packing was prepared by the immobilization of Cibacron Blue F3GA as dye ligands on the base of monodispersed, 3.0 microm 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 F3 GA and lysozyme (Lys) was 5.26 x 10(-5) mol/L which was evaluated by frontal chromatography. Purifications of Lys from egg white and bovine serum albumin (BSA) from 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 5% and 92%, respectively.