Polymer Brush-Modified Magnetic Nanoparticles for His-Tagged Protein Purification

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• 作者：Fei Xu ; James H. Geiger ; Gregory L. Baker ; Merlin L. Bruening
• 刊名：Langmuir
• 出版年：2011
• 出版时间：March 15, 2011
• 年：2011
• 卷：27
• 期：6
• 页码：3106-3112
• 全文大小：865K
• 年卷期：v.27,no.6(March 15, 2011)
• ISSN：1520-5827

文摘

Growth of poly(2-hydroxyethyl methacrylate) brushes on magnetic nanoparticles and subsequent brush functionalization with nitrilotriacetate-Ni²⁺ yield magnetic beads that selectively capture polyhistidine-tagged (His-tagged) protein directly from cell extracts. Transmission electron microscopy, Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric analysis, and magnetization measurements confirm and quantify the formation of the brushes on magnetic particles, and multilayer protein adsorption to these brushes results in binding capacities (220 mg BSA/g of beads and 245 mg His-tagged ubiquitin/g of beads) that are an order of magnitude greater than those of commercial magnetic beads. Moreover, the functionalized beads selectively capture His-tagged protein within 5 min. The high binding capacity and protein purity along with efficient protein capture in a short incubation time make brush-modified particles attractive for purification of recombinant proteins.