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采用反相悬浮包埋技术合成多分散的粒径在 5 0目～ 30 0目的磁性葡聚糖微球 (MDMS)。MDMS经环氧氯丙烷活化后 ,分别键合氨基乙酸、6 氨基己酸和乙二胺作为间隔臂 ,以碳二亚胺为偶联剂 ,分别偶联L 精氨酸甲酯、对氨基苯甲脒和胍基己酸配体 ,制备了 5种磁性亲和吸附剂。研究了分散介质及其粘度和密度、有机相和水相的体积比、表面活性剂的用量、搅拌速度等因素对微球制备的影响。将所制备的磁性亲和吸附剂应用于尿激酶粗品的纯化 ,并讨论了偶联试剂和配体对尿激酶纯化效果的影响。通过测定 ,尿激酶的活性回收率为 40 0 %～ 6 0 7%、纯化倍数为 14 9～ 32 8、吸附容量为 89mg/L～ 12 1mg/L。
Dispersion of magnetic dextran microspheres (MDMS) with particle size of 50 ~ 300 mesh was synthesized by reverse phase suspension technique. After activated by epichlorohydrin, MDMS was bonded with aminoacetic acid, 6-aminocaproic acid and ethylenediamine as spacer arms, carbodiimide as coupling agent, respectively coupled with L-arginine methyl ester and p-aminobenzene Formamidine and guanidinohexanoate ligands, five kinds of magnetic affinity adsorbents were prepared. The effects of dispersion medium, viscosity and density, volume ratio of organic and aqueous phases, the amount of surfactant and stirring speed on the preparation of microspheres were investigated. The prepared magnetic affinity adsorbent was applied to the purification of crude urokinase and the effect of coupling reagents and ligands on the purification of urokinase was discussed. The results showed that the recovery rate of urokinase was 40 0% ~ 60 7%, the purification fold was 14 9 ~ 32 8, and the adsorption capacity was 89 mg / L ~ 12 1 mg / L.