论文部分内容阅读
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.
The magnetic nanoparticle Fe3O4 magnetic core was synthesized by chemical coprecipitation, and the homemade magnetic core was coated with chitosan as the coating material. The magnetic micro-sphere with chitosan core-shell structure was prepared by emulsion cross-linking method. A new type of affinity magnetic microspheres was obtained by coupling heparin with ligands, and the particle size, morphology, structure and magnetic responsiveness of the microspheres were characterized by SEM, FT-IR and XRD. Affinity magnetic microspheres on thrombin separation and purification performance, and compared with the traditional DEAE ion exchange chromatography results show that the resulting affinity magnetic microspheres with a narrower particle size distribution, shape regular, particle size at 50nm , And the specific activity of 1879.71U / mg was obtained by one step adsorption-purification of thrombin, with the yield of 85% and the purification multiple of 11.057, while the yield of traditional column chromatography was 72% and the purification fold was only 5.33. Glycans affinity magnetic microspheres, magnetic separation technology and applied to the separation and purification of thrombin, get better results, which will have some reference value for the purification and production of thrombin.