高效 Ni-NTA/PAA 磁珠对 His 标记蛋白具有特异性分离。
High-efficiency Ni-NTA/PAA magnetic beads with specific separation on His-tagged protein.
机构信息
Hubei Key Laboratory of Industrial Microbiology, Hubei University of Technology, Wuhan 430068, People's Republic of China.
出版信息
IET Nanobiotechnol. 2020 Feb;14(1):67-72. doi: 10.1049/iet-nbt.2019.0271.
Abstract
To effective capture and universal enrichment of His-tagged protein, polyacrylic acid (PAA) brushes were used to encapsulate FeO nanoparticles, connect NTA, and Ni to prepare magnetic beads. These materials provide many advantages, such as excellent stability, tuneable particle size, and a surface for further functionalisation with biomolecules. His-tagged green fluorescence protein (GFP) was separated efficiently, and the binding capacity of FeO/MPS@PAA/NTA-Ni was 93.4 mg/g. Compared with High-Affinity Ni-NTA Resin and Ni-NTA Magnetic Agarose Beads, FeO/MPS@PAA/NTA-Ni nanocomposites exhibited higher separation efficiency and binding capacity towards His-tagged GFP. Moreover, the selectivity and recyclability of them for the target proteins were maintained well after six cycles. This study would widen the application of PAA in constructing multifunctional nanocomposites for biomedical fields.
摘要
为了有效捕获和普遍富集 His 标记蛋白，使用聚丙烯酸 (PAA) 刷来包裹 FeO 纳米颗粒，连接 NTA 和 Ni 以制备磁性珠。这些材料具有许多优点，如极好的稳定性、可调节的粒径以及可进一步与生物分子进行功能化的表面。高效地分离了 His 标记的绿色荧光蛋白 (GFP)，并且 FeO/MPS@PAA/NTA-Ni 的结合容量为 93.4mg/g。与高亲和力 Ni-NTA 树脂和 Ni-NTA 琼脂糖珠相比，FeO/MPS@PAA/NTA-Ni 纳米复合材料对 His 标记 GFP 具有更高的分离效率和结合容量。此外，在六次循环后，它们对目标蛋白的选择性和可回收性仍保持良好。这项研究将拓宽 PAA 在构建用于生物医学领域的多功能纳米复合材料中的应用。
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