染料结合2μm无孔硅胶亲和填料的制备与应用

在２μｍ无孔硅胶表面键合３ 氨丙基 三乙氧基硅烷（ＡＰＳ），并与三嗪染料活性蓝Ｆ３ＧＡ（ＣｉｂａｃｒｏｎＢｌｕｅＦ３ＧＡ，ＣＢ）反应，制得亲和色谱填料，并采用扫描电镜、元素分析、ｐＨ稳定性测试对此填料进行鉴定与表征。该填料具有良好的色谱性能，且对生物大分子有一定的亲和选择性，改变ｐＨ值及离子强度对溶菌酶的结合量有明显影响，可用于分离卵清蛋白（Ｏｖａｌ）和溶菌酶（Ｌｙｓ），且对α ，β ，γ 球蛋白有不同的亲和作用，并可从鸡蛋清中制备少量溶菌酶。

Cibacron blue F3GA-attached 2 microns non-porous monodisperse silicas for affinity chromatography]

Non-porous monodisperse silica (NPS), 2 microns in diameter, was modified with 3-aminopropyltriethoxysilane for immobilization of Cibacron Blue F3GA (CB), a packing of NPS-ACB for affinity chromatography was obtained. Up to 2 mg of CB could be attached to 1 mL of NPS beads. There was no obvious leakage of dye from NPS-ACB. Oval was not retained by the column, while Lys was specifically adsorbed. The adsorption of Lys varied with pH values and ionic strengths. In addition, alpha-globulin could not be retained by the packing, while beta- and gamma-globulin could be adsorbed on the column. gamma-Globulin was able to be eluted by 20% 1,6-hexanediol and 1 mol/L KCl, while beta-globulin was not able to be eluted by the same eluent. The difference in affinity interaction could be used to separate the three globulins. Furthermore, the column could be used for separation and preparation of Lys from hen egg white. The chromatograms of Lys on non-porous silica diethylamine column (NPS-DEA) showed that retention time of one peak of the crude Lys prepared was in accordance with Lys's, so it could be said that NPS-ACB column can be used for preparation in a small scale.