Magnetic cell separation using nano‐sized bacterial magnetic particles with reconstructed magnetosome membrane
Magnetic cell separation using nano‐sized bacterial magnetic particles with reconstructed magnetosome membrane
复制标题
使用具有重建磁小体膜的纳米细菌磁性颗粒进行磁性细胞分离
DOI：
10.1002/bit.21912
复制
发表时间：
2008
影响因子：
3.8
通讯作者：
T. Matsunaga
中科院分区：
文献类型：
--
作者：
T. Yoshino;Hisashi Hirabe;Masayuki Takahashi;Motoki Kuhara;H. Takeyama;T. Matsunaga
由趋磁细菌产生的磁性纳米颗粒，即细菌磁性颗粒（BacMPs），表面覆盖有脂质双层膜（磁小体膜），可用于从异质混合物中分离特定的靶细胞，因为它们易于被磁体操纵，并且易于通过基因工程在其表面展示功能蛋白。尽管具有独特的和有价值的特性，BacMPs的潜在的毒性分离的细胞尚未被详细描述。在这里，开发了一种新技术，用于重建表达蛋白A的BacMP（蛋白A-BacMP）的磁小体膜，以减少细胞毒性，然后将新开发的纳米材料用于磁性细胞分离。磁小体膜重构蛋白A-BacMP的开发基于Mms 13锚蛋白的特性，其与BacMP的磁铁矿表面强烈结合。用去污剂处理蛋白A-BacMP去除污染蛋白，但不影响Mms 13-蛋白A融合蛋白的保留。然后用磷脂酰胆碱重建颗粒表面。含有重建的磁小体膜的蛋白A-BacMP保持可分散性并保留了抑制抗体的能力。此外，它们含有很少的膜表面蛋白和内毒素，这在未处理的蛋白A-BacMP上观察到。使用磁小体膜重构蛋白A-BacMP实现了外周血单核细胞和B-淋巴细胞的高纯度磁性分离。Biotechnol. Bioeng. 2008;101：470-477.© 2008 Wiley Periodicals，Inc.
Magnetic nanoparticles produced by magnetotactic bacterium, bacterial magnetic particles (BacMPs), covered with a lipid bilayer membrane (magnetosome membrane) can be used to separate specific target cells from heterogeneous mixtures because they are easily manipulated by magnets and it is easy to display functional proteins on their surface via genetic engineering. Despite possessing unique and valuable characteristics, the potential toxicity of BacMPs to the separated cells has not been characterized in detail. Here, a novel technique was developed for the reconstruction of magnetosome membrane of BacMPs expressing protein A (protein A‐BacMPs) to reduce cytotoxicity and the newly developed nanomaterial was then used for magnetic cell separation. The development of the magnetosome membrane‐reconstructed protein A‐BacMP was based on the characteristics of the Mms13 anchor protein, which strongly binds to the magnetite surface of BacMPs. Treatment of protein A‐BacMPs with detergents removed contaminating proteins but did not affect retention of Mms13‐protein A fusion proteins. The particle surfaces were then reconstructed with phosphatidylcholine. The protein A‐BacMPs containing reconstructed magnetosome membranes remained dispersible and retained the ability to immobilize antibody. In addition, they contained few membrane surface proteins and endotoxins, which were observed on non‐treated protein A‐BacMPs. Magnetic separation of monocytes and B‐lymphocytes from the peripheral blood was achieved with high purity using magnetosome membrane‐reconstructed protein A‐BacMPs. Biotechnol. Bioeng. 2008;101: 470–477. © 2008 Wiley Periodicals, Inc.
影响因子：
15
作者：
Uchida, Masaki;Flenniken, Michelle L.;Douglas, Trevor
通讯作者：
Douglas, Trevor