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非离子型、温敏性 DEA/DMA 纳米凝胶:合成、表征及其在微芯片电泳中用于 DNA 分离的应用。

Non-ionic, thermo-responsive DEA/DMA nanogels: synthesis, characterization, and use for DNA separations by microchip electrophoresis.

机构信息

Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL 60208, USA.

出版信息

J Colloid Interface Sci. 2011 May 15;357(2):345-53. doi: 10.1016/j.jcis.2011.01.079. Epub 2011 Feb 1.

DOI:10.1016/j.jcis.2011.01.079
PMID:21392778
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3086585/
Abstract

Thermo-responsive polymer "nanogels" (crosslinked hydrogel particles with sub-100 nm diameters) are intriguing for many potential applications in biotechnology and medicine. There have been relatively few reports of electrostatically neutral, thermosensitive nanogels comprising a high fraction of hydrophilic co-monomer. Here we demonstrate the syntheses and characterization of novel, non-ionic nanogels based on random N,N-diethylacrylamide (DEA)/N,N-dimethylacrylamide (DMA) copolymers, made by free-radical, surfactant-free dispersion polymerization. The volume-phase transition temperatures of these DEA/DMA nanogels are strongly affected by co-monomer composition, providing a way to "tune" the phase transition temperature of these non-ionic nanogels. While DEA nanogels (comprising no DMA) can be obtained at 70 °C by standard emulsion precipitation, DEA/DMA random co-polymer nanogels can be obtained only in a particular range of temperatures, above the initial phase transition temperature and below the critical precipitation temperature of the DEA/DMA copolymer, controlled by co-monomer composition. Increasing percentages of DMA in the nanogels raises the phase transition temperature, and attenuates and broadens it as well. We find that concentrated DEA/DMA nanogel dispersions are optically clear at room temperature. This good optical clarity was exploited for their use in a novel DNA sieving matrix for microfluidic chip electrophoresis. An ultrafast, high-efficiency dsDNA separation was achieved in less than 120 s for dsDNA ranging from 75 bp to 15,000 bp.

摘要

温敏聚合物“纳米凝胶”(亚 100nm 直径的交联水凝胶颗粒)在生物技术和医学的许多潜在应用中都很有吸引力。相对较少有报道关于静电中性、包含高比例亲水性共聚单体的热敏纳米凝胶。在这里,我们展示了基于无规 N,N-二乙基丙烯酰胺(DEA)/N,N-二甲基丙烯酰胺(DMA)共聚物的新型非离子纳米凝胶的合成和表征,这些共聚物是通过自由基、无表面活性剂的分散聚合制备的。这些 DEA/DMA 纳米凝胶的体积相转变温度强烈受到共聚单体组成的影响,为“调谐”这些非离子纳米凝胶的相转变温度提供了一种方法。虽然 DEA 纳米凝胶(不包含 DMA)可以通过标准乳液沉淀在 70°C 下获得,但 DEA/DMA 无规共聚物纳米凝胶只能在特定的温度范围内获得,该温度范围高于初始相转变温度,低于 DEA/DMA 共聚物的临界沉淀温度,由共聚单体组成控制。纳米凝胶中 DMA 的百分比增加会提高相转变温度,并使其变窄变宽。我们发现,在室温下,浓度较高的 DEA/DMA 纳米凝胶分散体具有良好的光学透明度。这种良好的光学透明度被用于微流控芯片电泳的新型 DNA 筛分基质。对于从 75bp 到 15000bp 的 dsDNA,实现了不到 120s 的超快、高效的 dsDNA 分离。