用微悬浮共聚合技术实现了二乙烯基苯与醋酸乙烯酯单体对亲油性Fe3O4纳米粒子的充分包裹, 制备出超顺磁性的聚合物微球, 研究了分离人血清抗体的特异性.结果表明, 聚合物微球的粒径为0.6--20μm, 磁性微球的含量高于24%,饱和磁化强度为16.7 emu/g. 通过充分的水解反应, 在微球表面形成丰富的羟基,并通过Michael加成反应, 在磁性微球表面偶联嗜硫基团(乙烯基砜和巯基乙醇).用嗜硫性磁性微球直接从人血清中分离免疫球蛋白G, 表现出明显的特异性吸附行为.在分离过程中, 吸附行为表现出明显的盐依赖性, 硫酸钠能有效促进其特异性吸附能力.这种吸附行为对温度比较敏感, 在低温条件下吸附能力和分离效率皆降低.利用磁性微球技术的抗体纯化过程, 工艺条件简单, 分离条件温和, 抗体纯度较高,在规模化分离抗体方面表现出较大的潜力.
The paramagnetic polymer particles with the size range of 0.6–20 μm were synthesized by microsuspension copolymerization with vinyl acetate(VAc) and divinylbenzene(DVB) in the presence of modified magnetite nanoparticles. Evident paramagnetic behavior was observed and saturation magnetization reached 16.7 emu·g−1 at room temperature. These magnetic microspheres were first activated by divinylsulfone, and then modified with mercaptoethanol. By using these thiophilic magnetic microspheres, immunoglobulin G (IgG) could be directly isolated from human serum. This thiophilic magnetic adsorbent performed an evident salt-dependent adsorption behavior for IgG. Due to their strong specificity towards IgG under high salt concentration, the absorbed antibodies could be extracted with high purity. The main factors to influence the isolation, such as the ion types, the ion concentration and the temperature, were also investigated. The technology has great potential to purify antibodies from human serum.
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