染料膜亲和色谱法中膜堆的制备及应用  

Preparation and Application of the Cartridges. of Dye Membrane Affinity Chromatography

作　　者：郭为[1] 商振华[1] 于亿年[1] 周良模[1] 

机构地区：[1]中国科学院大连化学物理研究所

出　　处：《色谱》1996年第3期168-171,共4页Chinese Journal of Chromatography

基　　金：国家自然科学基金

摘　　要：将染料亲和配基偶联于大孔纤维素膜上，所得亲和膜用胶粘法制成亲和膜堆，膜堆的通透性远优于通常的亲和色谱柱。装有蓝色和红色亲和膜的膜堆可分别用于人血清白蛋白和碱性磷酸酯酶的分离纯化，其中碱性磷酸酯酶可在一步操作后纯化４０倍。n attractive membrane cartridge using macropore cellulose membrane as stationary phase and newly designed shell was prepared. The membrane capacity of the cartridge was adjustable depending on the membrane bundlings with different height. The distribution of mobile phase on the membrane surface was uniformed by installing distribution plates in front of and behind the medium. To solve the problem of radial leakage, a special bonding method was adopted, in which the circumference with a width of 1 mm on the edge of each individual membrane was saturated with an elastic glue. This method can help the sealing and enhancing the permeability of the cartridge. The pressure drop across the cartridge increased linearly according to the flow rate, and the flux of cartridge containing 80 sheets of membrane can reach 5. 0 mL/min with a pressure drop of 0.1 MPa.Triazine dyes, Cibacron Blue F3GA and Active Red K2BP, were immobilized as affinity ligands. The membrane capacity for dye immobilization can be much improved by alkaline treatment. UP to 90 mg of Active Red K2BP or 100 mg Cibecron Blue F3GA can be coupled onto 1 g of membrane matrix under the optimum condition. The cleavage of dye ligand from matrix was investigated and no leakage was found when the PH value of the eluent was more than 3. Membrane cartridges containing blue and red membrane were also prepared. Human serum albumin and alkaline phosphatase were purified on blue and red affinity membrane cartridges respectively. For alkaline phosphatase purification, a 40-fold enrichment and a 600% activity recovery were achieved.

关 键 词：膜亲和色谱 磷酸酯酶 人血清 白蛋白 

分 类 号：O652.63[理学—分析化学] Q512.1[理学—化学]