二次乳化法(W/O)/W制备蛋白质流通色谱固定相的研究

以甲基丙烯酸缩水甘油酯为单体,二甲基丙烯酸乙二醇酯为交联剂,甲苯与正庚烷为有机致孔剂,20%的甘油水溶液为内水相(超孔致孔剂),利用二次乳化法制备了(W/O)/W乳液,通过紫外光引发悬浮聚合生成两类孔型高分子微球(BiPB).BiPB孔径为双峰分布,范围分别在10~100nm和1000~7300nm之间;而其体积平均粒径、比表面积、湿密度、静态吸附容量与不含超孔的微孔介质(MiPB)接近.修饰相同二乙胺密度的BiPB和MiPB介质虽在较低流速(5cm/min)下有相近的动态吸附容量,但在高流速(40cm/min)下BiPB的动态吸附容量约为MiPB的动态吸附容量的3倍,表明BiPB介质的超孔结构对孔内传质的强化作用,因而其更适合于高速的蛋白质色谱分离.

PREPARATION OF A BIPOROUS POLYMERIC ADSORBENT BY DOUBLE EMULSIFICATION FOR FLOWTHROUGH CHROMATOGRAPHY OF PROTEINS

This paper reports a technique to fabricate the stationary phase for high-speed protein flowthrough chromatography.Using glycerol aqueous solution (20 wt%) and n-heptane/toluene organic solvent as the porogenic agents,a novel type of biporous matrix (BiPB) has been prepared by ultraviolet photo-polymerizing monomers (glycidyl methacrylate and ethylene glycol dimethacrylate) by a (W/O)/W emulsification technique.It was found that there were two types of pores (micropores and superpores) in BiPB,but there was only micropores in the microporous beads (MiPB) created by O/W emulsification.The pores of BiPB were in the range of 10～100 nm and 1000～7300 nm,whereas those of MiPB were mainly 10～100 nm in size.The specific surface area,hydrated density and particle size of BiPB were 41.9 m~2/g,1.08 g/mL and 42.8 μm,respectively,similar to those of MiPB (52.1 m~2/g,1.11 g/mL and 42.4 μm).The static adsorption capacity of BiPB for bovine serum albumin (BSA) was also similar to that of MiPB.At a low flow rate (5 cm/min),the dynamic adsorption capacity (DAC) of BiPB for BSA was close to that of MiPB.At high flow rate (40 cm/min),however,the DAC of BiPB was about three times as high as that of MiPB.The results indicate that BiPB columns were promising for high-speed protein chromatography.