用壳聚糖亲和磁性微球纯化血浆凝血酶的研究  

The Study of Purifying Plasma Thrombin Using Chitosan Affinity Magnetic Microspheres

作　　者：李梅基[1,2] 李昭华[2] 胡建成[1] 董先智[2] 

机构地区：[1]兰州大学生命科学学院,兰州730000 [2]中国科学院生物物理研究所,北京100101

出　　处：《生物化学与生物物理进展》2010年第4期433-440,共8页Progress In Biochemistry and Biophysics

摘　　要：通过化学共沉淀法合成纳米粒子Fe3O4磁核,以壳聚糖为包裹材料包被自制的磁核,采用乳化交联法制备了具有核-壳结构的磁性高分子微球-壳聚糖磁性微球,并偶联肝素配基得到了一种新型亲和磁性微球,应用SEM、FT-IR、XRD等对微球的粒径、形貌、结构和磁响应性进行了表征.考察了该亲和磁性微球对凝血酶的分离纯化性能,并与传统的DEAE离子交换色谱法进行了比较.结果表明,所得亲和磁性微球具有较窄的粒径分布、形状规整,粒径在50nm左右.对凝血酶一步吸附纯化获得了比活为1879.71U/mg的酶,得率85%,纯化倍数11.057,而传统柱层析法得率为72%,纯化倍数仅为5.33.制备了壳聚糖亲和磁性微球,并将磁分离技术应用于凝血酶的分离纯化,得到了较好的效果,这将对于凝血酶的纯化及生产具有一定参考价值.Nanometer Fe3O4 particle was synthesized by coprecipitation, magnetic polymeric microspheres- magnetic chitosan microspheres, containing a magnetic core and a polymer shell, were prepared by the suspension cross-linking technique. Novel affinity magnetic microspheres were obtained after immobilizing heparin onto the nanoparticles. The size, morphology, structure and magnetic response properties were studied by means of SEM, FT-IR and XRD and so on. The novel affinity magnetic microspheres′ properties for thrombin were examined by its adsorption and purification, and compared with the traditional means of column chromatography— DEAE-Sepharose Fast Flow. The results showed the affinity magnetic microspheres had a spherical appearance, narrow distribution of diameter, the particle diameter is about 50 nm, and one-step purification of thrombin using it the specific activity was 1 879.7 U/mg, recovery rate was 85% and purification fold was 11.057. Whereas, purification of thrombin by column chromatography, the product exhibits an activity of 909.84 U/mg, recovery rate of 72% and purification fold of 5.33. In conclusion, chitosan nanometer affinity magnetic microspheres was prepared and magnetic separation techniques was applied for purification of thrombin, the results is better than that from traditional methods and these results have some references values in the production and subsequent study on the research and development.

关 键 词：亲和磁性微球 壳聚糖 凝血酶 磁分离 纯化 

分 类 号：Q658[生物学—生物物理学]