二次乳化法(W/O)/W制备蛋白质流通色谱固定相的研究

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

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，因而其更适合千高谏的蛋白后仁．

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.