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通过包裹有次氮基三乙酸(NTA)-磷脂胶束的磁性纳米颗粒固定组氨酸标签蛋白。

Immobilization of histidine-tagged proteins by magnetic nanoparticles encapsulated with nitrilotriacetic acid (NTA)-phospholipids micelle.

作者信息

Lim Yong Taik, Lee Kun Yeong, Lee Kwangyeol, Chung Bong Hyun

机构信息

Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, 52 Eoeun-dong, Yuseong-gu, Daejeon 305-333, Republic of Korea.

出版信息

Biochem Biophys Res Commun. 2006 Jun 9;344(3):926-30. doi: 10.1016/j.bbrc.2006.03.209. Epub 2006 Apr 19.

Abstract

We described the development of functionalized magnetic nanoparticles (MNPs) with PEG-modification, a phospholipids micelle coating, and their use in manipulating histidine-tagged proteins. Highly monodisperse MNPs were synthesized in an organic solvent and could be phase-transferred into an aqueous solution by encapsulating the nanoparticles with a phospholipids micelle. The phospholipids micelle coating rendered the nanoparticles highly water-soluble, and the functional groups of the phospholipids coating allowed for the bioconjugation of various moieties, such as fluorescent molecules and engineered proteins. Functionalized phospholipids, such as nitrilotriacetic acid (NTA)-phospholipids, caused the MNPs to bind and allowed for manipulation of histidine-tagged proteins. Due to their high surface/volume ratio, the MNPs showed better performance (about 100 times higher) in immobilizing engineered proteins than conventional micrometer-sized beads. This demonstrates that MNPs coated with phospholipids micelle can be a versatile platform for the effective manipulation of various kinds of engineered proteins, which is very important in the field of proteomics. It is expected that a combination of MNPs with optical fluorescent molecules can find applications in bimodal (magnetic and optical) molecular imaging nanoprobes.

摘要

我们描述了具有聚乙二醇(PEG)修饰、磷脂胶束涂层的功能化磁性纳米颗粒(MNP)的开发及其在处理组氨酸标签蛋白中的应用。在有机溶剂中合成了高度单分散的MNP,通过用磷脂胶束包裹纳米颗粒可将其相转移到水溶液中。磷脂胶束涂层使纳米颗粒具有高度水溶性,并且磷脂涂层的官能团允许各种部分(如荧光分子和工程蛋白)进行生物共轭。功能化磷脂,如次氮基三乙酸(NTA)-磷脂,使MNP能够结合并处理组氨酸标签蛋白。由于其高表面/体积比,MNP在固定工程蛋白方面比传统的微米级珠子表现出更好的性能(约高100倍)。这表明涂有磷脂胶束的MNP可以成为有效处理各种工程蛋白的通用平台,这在蛋白质组学领域非常重要。预计MNP与光学荧光分子的组合可用于双模态(磁性和光学)分子成像纳米探针。

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