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利用疏水性磁珠从人血清中纯化抗体。

Using thiophilic magnetic beads in purification of antibodies from human serum.

机构信息

College of Material Science and Engineering, Huaqiao University, Quanzhou 362021, Fujian, China.

出版信息

Colloids Surf B Biointerfaces. 2010 Jan 1;75(1):342-8. doi: 10.1016/j.colsurfb.2009.09.007. Epub 2009 Sep 15.

Abstract

The rapid progress of biotechnology, immunology and molecular biology are requiring antibodies with higher purity and stronger activity. Therefore, the development of efficient technology, which is suitable for large-scale purification of antibodies at low cost, becomes much more urgent. In this study, we presented a novel method to isolate immunoglobulin G from human serum based on the utilization of thiophilic magnetic polymer beads. Micron-sized magnetic beads with paramagnetic properties were synthesized by microsuspension polymerization in the presence of modified magnetite. After the thiophilic ligand of 2-mercaptonicotinic acid was modified on the surface, these magnetic beads exhibited a strong specificity towards immunoglobulin G in a salt-independent manner. Then, antibodies could be directly isolated from human serum in batch-wise mode with the assistance of magnetic decantation. The purity of the isolated antibody exceeded 94%. Because isolation was performed in physiological conditions, bioactivity of the antibody was fully preserved (>99%). Prominent advantages of this method, such as strong specificity, rapid processing, mild conditions, conventional equipment and excellent reusability, make this non-chromatographic technology embody great potentialities to isolate the antibodies on a large scale.

摘要

生物技术、免疫学和分子生物学的快速发展,要求抗体具有更高的纯度和更强的活性。因此,开发高效的技术变得更加紧迫,这种技术应能够以低成本大规模地纯化抗体。在本研究中,我们提出了一种基于利用疏硫亲磁聚合物珠从人血清中分离免疫球蛋白 G 的新方法。在存在改性磁铁矿的情况下,通过微悬浮聚合合成了具有顺磁性的微米级磁性珠。在表面修饰了 2-巯基烟酸的疏硫配体后,这些磁性珠表现出了在盐非依赖性方式下对免疫球蛋白 G 的强特异性。然后,在磁沉降的辅助下,可以直接从人血清中批量分离抗体。分离得到的抗体的纯度超过 94%。由于分离是在生理条件下进行的,抗体的生物活性得到了充分保留(>99%)。该方法具有特异性强、处理速度快、条件温和、设备常规和可重复使用性好等显著优点,使这种非色谱技术在大规模分离抗体方面具有巨大的潜力。

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