离子交换与吸附

通过合成条件的改进,得到了一种性能更好的扩张床介质GMA-EDMA-Lipo-TiO2(GELT),其平均密度为1.25g/ml,孔容为0.564ml/g,平均孔径为15.6nm,比表面积为127.58m2/g,渗磨圆球率为95%。研究表明GELT在氢氧化钠溶液是稳定的。同时讨论了甲苯致孔剂对孔结构的影响及二氧化钛添加量对湿真密度的影响;将GELT与二乙胺偶联得到阴离子树脂(GELT-DEAE),在静态和动态条件下,GELT-DEAE吸附量分别88mgBSA/ml、65mgBSA/ml。将GELT-DEAE应用于扩张床直接从发酵液中分离复合干扰素,一步活性收率为75%,后经SPSepharoseFF离子交换层析和CellufastGel01凝胶层析进一步纯化。最后得到的成品其比活为6.82×108IU/mg,与商品干复津(Infergen)的比活相当,纯度为95%。

By improving the conditions of synthesis,a medium GMA-EDMA-Lipo-TiO2 (GELT) was obtained, with an average wet true density of 1.25mg/ml, pore volume 0.53ml/g, average pore diameter 15.6nm, specific surface area 127.6m2/g, and a sphericity after osmotic attrition of 95%. The obtained GELT was stable in NaOH solution. The effect of toluene as a porogenic agent and the amount of Lipo-TiO2 on the porous structure and wet true density of GELT was investigated. GELT coupled with diethylamine, formed an anion exchanger (GELT-DEAE), Its static and dynamic adsorption capacity in expanded bed were 88mgBSA/ml and 65mgBSA/ml, respectively. GELT-DEAE was used directly to separate IFN-Con-1 from broth in expanded bed mode with an activity yield of 75%, and then consecutively purified through a SP Sepharose FF column and a Cellufast Gel01 column, obtaining a final product with a purity of 95%.and a specific activity of 6.82×108 IU/mg, similar to infergen.