免疫磁性微球的制备及对IgG的分离

发布时间：2019-05-18 07:56

【摘要】： 免疫磁性微球(Immunomagnetic Microspheres,简称IMMS)是一种兼具磁性材料和高分子材料性能的新型功能材料,由于粒径小,比表面积大,可偶联的生物分子容量大,且能分散于体系中不易沉降,非常适合在生物体系中使用。免疫磁性分离法由于具有高效、快速、低毒、操作简单等优点,迄今已在细胞生物学、生物医学和生物工程等诸多领域展现了广阔的应用前景。本论文利用葡萄球菌蛋白A(Staphylococcal Protein A,简称SPA)对免疫球蛋白G(IgG)的特异性吸附反应及磁性微球的超顺磁性,从猪血清中分离IgG,其主要内容为: 采用化学沉淀法与球磨法制备Fe_3O_4磁性微球,通过探讨搅拌速度、表面活性剂加入量、球磨速度、球磨时间、球料比等各种因素对产物的粒径及磁性能的影响,及对Fe_3O_4磁性微球进行粒径、磁性能、铁含量等性能表征的研究。制得的产物具有较好的结构,粒径小于20nm,磁响应性强,具有超顺磁性,粒径分布均匀,分散性良好。采用反相悬浮包埋法制备SPA-Sepharose免疫磁性微球,对产物进行了结构,粒径、磁性能的表征,探讨了搅拌速度、表面活性剂、难溶助剂等因素对免疫磁性微球性能的影响,制取的产物粒径大约为5μm,表面包裹SPA,分散性较好,具有超顺磁性。试验结果表明,SPA免疫磁性微球对猪血清的最大吸附效率是70.12%,可反复使用29次。本实验为从猪血清中分离、纯化IgG的开辟了一条新的途径,为其产业化提供了一定的实验基础和理论依据。
[Abstract]:Immunommagnetic microspheres (Immunomagnetic Microspheres, (IMMS) is a new functional material with both magnetic and polymer properties. Because of its small particle size, large specific surface area, large coupling biomolecule capacity, and can not be dispersed in the system, it is not easy to settle. It is very suitable for use in biological systems. Immunomagnetism method has shown broad application prospects in many fields, such as cell biology, biomedicine, bioengineering and so on, because of its high efficiency, rapidity, low toxicity, simple operation and so on. In this paper, the specific adsorption reaction of Staphylococcus protein A (Staphylococcal Protein A, (SPA) for immunoglobulin G (IgG) and the superparamagnetism of magnetic microspheres were used to isolate IgG, from pig serum. The main contents are as follows: Fe_3O_4 magnetic microspheres were prepared by chemical precipitation method and ball milling method. The stirring speed, the amount of surfactants, the ball milling speed and the ball milling time were discussed. The effects of ball / material ratio on the particle size and magnetic properties of the products, as well as the characterization of the particle size, magnetic properties and iron content of Fe_3O_4 magnetic microspheres were studied. The prepared product has a good structure, the particle size is less than 20 nm, the magnetic response is strong, it has superparamagnetism, the particle size distribution is uniform and the dispersion is good. SPA-Sepharose immunommagnetic microspheres were prepared by inverse suspension embedding method. The structure, particle size and magnetic properties of the products were characterized. The effects of stirring speed, surfactants and insoluble auxiliaries on the properties of immunommagnetic microspheres were discussed. The particle size of the product is about 5 渭 m, and the surface coated SPA, has good dispersion and superparamagnetism. The results showed that the maximum adsorption efficiency of SPA immunommagnetic microspheres for pig serum was 70.12% and could be reused 29 times. This experiment opens up a new way to isolate and purify IgG from pig serum, and provides a certain experimental and theoretical basis for its industrialization.
【学位授予单位】：西华大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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