Aptamer-Based Downstream Processing of His-Tagged Proteins Utilizing Magnetic Beads
Aptamer-Based Downstream Processing of His-Tagged Proteins Utilizing Magnetic Beads
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DOI：
10.1002/bit.23191
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发表时间：
2011-10-01
影响因子：
3.8
通讯作者：
Scheper, Thomas
中科院分区：
文献类型：
--
作者：
Koekpinar, Oeznur;Walter, Johanna-Gabriela;Scheper, Thomas
适体是合成的基于核酸的高亲和力配体，其能够通过分子识别捕获其相应的靶标。在这里，开发了基于适体的His标记蛋白的亲和纯化。将两种不同的针对His标签的适体共价固定在磁珠上。将所得的适体修饰的磁珠进行表征，并成功地应用于从复合物E中纯化不同的His标记的蛋白。大肠杆菌细胞裂解物。在一个单一的纯化步骤中实现了与传统的固定化金属亲和色谱相当的纯化效果。此外，我们已经研究了再生和重复使用适体修饰的磁珠的可能性，并显示了它们在6个月内的长期稳定性。Biotechnol. Bioeng. 2011; 108：2371-2379. (C)2011 Wiley Periodicals，Inc.
Aptamers are synthetic nucleic acid-based high affinity ligands that are able to capture their corresponding target via molecular recognition. Here, aptamer-based affinity purification for His-tagged proteins was developed. Two different aptamers directed against the His-tag were immobilized on magnetic beads covalently. The resulting aptamer-modified magnetic beads were characterized and successfully applied for purification of different His-tagged proteins from complex E. coli cell lysates. Purification effects comparable to conventional immobilized metal affinity chromatography were achieved in one single purification step. Moreover, we have investigated the possibility to regenerate and reuse the aptamer-modified magnetic beads and have shown their long-term stability over a period of 6 months. Biotechnol. Bioeng. 2011; 108: 2371-2379. (C) 2011 Wiley Periodicals, Inc.