Protein A亲和吸附磁珠的制备与抗体的快速纯化
首发时间:2020-03-09
摘要:以琼脂糖、Fe3O4粉末为原料,采用反向悬浮乳化法,制备出琼脂糖磁性复合微球,并偶联蛋白A功能配基,制备了Protein A亲和层析介质。采用扫描电镜、VSM、振动样品磁强计等方法对微球进行表征,结果表明:微球球形度良好、粒径分布均匀、四氧化三铁包埋率高、饱和磁化强度高、矫顽力低,具有较好的顺磁性。筛选磁珠粒径分别分布在25-38 μm、38-50 μm、50-70 μm、75-100 μm的Protein A磁珠微球,以不同粒径分布的磁珠做动力学吸附测试,结果表明粒径分布较小的微球在较短的时间内体现出更好的吸附效果。采用自制的Protein A亲和层析介质对小鼠血清进行吸附应用研究。结果显示:经吸附洗脱的IgG纯度能达到95%以上,平均粒径在31.0 μm磁珠适合用于快速纯化工艺。
关键词: 琼脂糖磁珠 Protein A 抗体 吸附 快速纯化。
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Protein A affinity Magnetic Beads for Antibody Fast Purification
Abstract:Agarose magnetic beads were prepared by the method of water-in-oil suspension with mixing of agarose and ferroferric oxide powder. Protein A was coupled on to this beads as a kind of Protein A affinity chromatography medium. The magnetic beads were characterized by SEM, VSM, et al. and the results showed that the microspheres had good sphericity, narrow size distribution, high embedding rate of ferroferric oxide, high magnetic saturation value, low coercivity and better paramagnetism. Magnetic beads with different size distribution and average diameter were screened to investigate the adsorption performance. The kinetic adsorption was determined and the results showed that the smaller beads exhibit better adsorption in the short time. Protein A magnetic beads were applied in purification of IgG from mouse serum. The results showed that the purity of eluted IgG reach more than 95%, and the magnetic beads with an average diameter of 31.0 μm are suitable used in fast purification.
Keywords: Agarose magnetic beads Protein A IgG Adsorption Fast purification
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Protein A亲和吸附磁珠的制备与抗体的快速纯化
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