發(fā)布時(shí)間：2019-05-18 07:56

【摘要】： 免疫磁性微球(Immunomagnetic Microspheres,簡(jiǎn)稱IMMS)是一種兼具磁性材料和高分子材料性能的新型功能材料,由于粒徑小,比表面積大,可偶聯(lián)的生物分子容量大,且能分散于體系中不易沉降,非常適合在生物體系中使用。免疫磁性分離法由于具有高效、快速、低毒、操作簡(jiǎn)單等優(yōu)點(diǎn),迄今已在細(xì)胞生物學(xué)、生物醫(yī)學(xué)和生物工程等諸多領(lǐng)域展現(xiàn)了廣闊的應(yīng)用前景。 本論文利用葡萄球菌蛋白A(Staphylococcal Protein A,簡(jiǎn)稱SPA)對(duì)免疫球蛋白G(IgG)的特異性吸附反應(yīng)及磁性微球的超順磁性,從豬血清中分離IgG,其主要內(nèi)容為: 采用化學(xué)沉淀法與球磨法制備Fe_3O_4磁性微球,通過(guò)探討攪拌速度、表面活性劑加入量、球磨速度、球磨時(shí)間、球料比等各種因素對(duì)產(chǎn)物的粒徑及磁性能的影響,及對(duì)Fe_3O_4磁性微球進(jìn)行粒徑、磁性能、鐵含量等性能表征的研究。制得的產(chǎn)物具有較好的結(jié)構(gòu),粒徑小于20nm,磁響應(yīng)性強(qiáng),具有超順磁性,粒徑分布均勻,分散性良好。 采用反相懸浮包埋法制備SPA-Sepharose免疫磁性微球,對(duì)產(chǎn)物進(jìn)行了結(jié)構(gòu),粒徑、磁性能的表征,探討了攪拌速度、表面活性劑、難溶助劑等因素對(duì)免疫磁性微球性能的影響,制取的產(chǎn)物粒徑大約為5μm,表面包裹SPA,分散性較好,具有超順磁性。 試驗(yàn)結(jié)果表明,SPA免疫磁性微球?qū)ωi血清的最大吸附效率是70.12%,可反復(fù)使用29次。本實(shí)驗(yàn)為從豬血清中分離、純化IgG的開(kāi)辟了一條新的途徑,為其產(chǎn)業(yè)化提供了一定的實(shí)驗(yàn)基礎(chǔ)和理論依據(jù)。
[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.
【學(xué)位授予單位】：西華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

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本文編號(hào)：2479812