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不同表面涂层的 FeO 基磁性纳米吸附剂在核酸提取中的应用比较评价。

Comparative Evaluation of Different Surface Coatings of FeO-Based Magnetic Nano Sorbent for Applications in the Nucleic Acids Extraction.

机构信息

Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Stanisława Noakowskiego 3, 00-664 Warsaw, Poland.

Doctoral School No. 1, Warsaw University of Technology, Plac Politechniki 1, 00-661 Warsaw, Poland.

出版信息

Int J Mol Sci. 2022 Aug 9;23(16):8860. doi: 10.3390/ijms23168860.

Abstract

Nucleic acid extraction and purification are crucial steps in sample preparation for multiple diagnostic procedures. Routine methodologies of DNA isolation require benchtop equipment (e.g., centrifuges) and labor-intensive steps. Magnetic nanoparticles (MNPs) as solid-phase sorbents could simplify this procedure. A wide range of surface coatings employs various molecular interactions between dsDNA and magnetic nano-sorbents. However, a reliable, comparative evaluation of their performance is complex. In this work, selected FeO modifications, i.e., polyethyleneimine, gold, silica, and graphene derivatives, were comprehensively evaluated for applications in dsDNA extraction. A family of single batch nanoparticles was compared in terms of morphology (STEM), composition (ICP-MS/MS and elemental analysis), surface coating (UV-Vis, TGA, FTIR), and MNP charge (ζ-potential). ICP-MS/MS was also used to unify MNPs concentration allowing a reliable assessment of individual coatings on DNA extraction. Moreover, studies on adsorption medium (monovalent vs. divalent ions) and extraction buffer composition were carried out. As a result, essential relationships between nanoparticle coatings and DNA adsorption efficiencies have been noticed. FeO@PEI MNPs turned out to be the most efficient nano sorbents. The optimized composition of the extraction buffer (medium containing 0.1 mM EDTA) helped avoid problems with Fe stripping, which improved the validity of the spectroscopic determination of DNA recovery.

摘要

核酸提取和纯化是多种诊断程序样品制备的关键步骤。常规的 DNA 分离方法需要台式设备(如离心机)和劳动密集型步骤。磁性纳米颗粒(MNPs)作为固相吸附剂可以简化此过程。广泛的表面涂层采用 dsDNA 和磁性纳米吸附剂之间的各种分子相互作用。然而,对其性能进行可靠、比较评估是复杂的。在这项工作中,对选定的 FeO 修饰物,即聚乙烯亚胺、金、硅和石墨烯衍生物,进行了全面评估,以应用于 dsDNA 提取。一组单批次纳米颗粒在形态(STEM)、组成(ICP-MS/MS 和元素分析)、表面涂层(UV-Vis、TGA、FTIR)和 MNP 电荷(ζ-电位)方面进行了比较。ICP-MS/MS 还用于统一 MNPs 浓度,从而可以可靠地评估 DNA 提取过程中各个涂层的性能。此外,还研究了吸附介质(单价离子与二价离子)和提取缓冲液组成。结果,注意到纳米颗粒涂层与 DNA 吸附效率之间存在重要关系。FeO@PEI MNPs 是最有效的纳米吸附剂。优化的提取缓冲液组成(含有 0.1 mM EDTA 的介质)有助于避免 Fe 剥离问题,从而提高了 DNA 回收的光谱测定的有效性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7240/9408759/ee0b57eb6eee/ijms-23-08860-g001.jpg

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