CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China.
University of Chinese Academy of Sciences, Beijing, P. R. China.
Electrophoresis. 2019 Aug;40(16-17):2135-2141. doi: 10.1002/elps.201900061. Epub 2019 Apr 30.
The binding coverage of aptamer was an important restricted factor for aptamer-based affinity enrichment strategy for capturing target molecules. Herein, we designed and prepared aptamer functionalized graphene oxide based nanocomposites (GO/NH -NTA/Fe O /PEI/Au), and the coverage density of aptamer was high to 33.1 nmol/mg. The high aptamer coverage density was contributed to the large surface area of graphene oxide. The successive modification of Nα,Nα-Bis(carboxymethyl)-L-lysine, magnetic nanoparticles, polyethylenimine, and Au nanoparticles ensured the histone purification with fast speed and high purity. Histones could be captured rapidly and specifically from nucleoproteins by our aptamer based purification strategy, while traditional acid-extraction could not specifically enrich histones. Compared with traditional acid-extraction method, rapid and efficient discovery of histones and their post-translational modifications, such as several kinds of methylation at H3.1K9 and H3.1K27, were achieved confidently. It demonstrated that our aptamer functionalized magnetic graphene oxide nanocomposites have a great potential for histone analysis.
适体的结合覆盖度是适体亲和富集策略用于捕获靶分子的一个重要限制因素。在此，我们设计并制备了适体功能化的基于氧化石墨烯的纳米复合材料（GO/NH-NTA/Fe3O4/PEI/Au），其适体的覆盖密度高达 33.1 nmol/mg。高的适体覆盖密度归因于氧化石墨烯的大表面积。Nα,Nα-双（羧甲基）-L-赖氨酸、磁性纳米粒子、聚乙烯亚胺和金纳米粒子的连续修饰确保了组蛋白的快速高纯度纯化。通过我们基于适体的纯化策略，组蛋白可以从核蛋白中快速特异性地捕获，而传统的酸提取则不能特异性地富集组蛋白。与传统的酸提取方法相比， confidently 实现了组蛋白及其翻译后修饰（如 H3.1K9 和 H3.1K27 上的几种甲基化）的快速高效发现。这表明我们的适体功能化磁性氧化石墨烯纳米复合材料在组蛋白分析方面具有很大的潜力。
