Milano Giovanna, Musumeci Domenica, Gaglione Maria, Messere Anna
Department of Environmental Sciences, Second University of Naples, via Vivaldi 43, 81100 Caserta, Italy.
Mol Biosyst. 2010 Mar;6(3):553-61. doi: 10.1039/b915680a. Epub 2009 Dec 11.
In this paper we report an alternative approach to synthesize PNA and DNA magnetic nanoconjugates. Chemical modifications were introduced on the 130 nm dextran-magnetite particles to obtain poly-functionalized particles containing reversible bonds sensitive to the cellular environment and suitable for the direct introduction of unmodified oligomers. Due to the polyvalent nature of the nanoparticles, when the complementary PNA and DNA nanoconjugates were mixed together, the resulting duplex structures bring to a nanoparticle assembly driven by W-C base pairs. The formation of the nanoparticle assembly was investigated by optical spectroscopy (UV, FTIR), scanning and transmission electron microscopies and by the analysis of the macroscopic behaviour of the nanoparticle-conjugates in aqueous solution with and without magnetic field application. Furthermore, serum stability assays revealed an increased enzymatic resistance in FCS of the PNA/DNA nanoconjugate duplex with respect to the unconjugated duplex. The described nanosystem could be extended to other duplex structures, possibly involving aptameric sequences of biomedical relevance, and could be very useful in order to obtain high local concentration at the target site of both the duplex and the magnetic nanoparticles in biotechnological applications.
在本文中,我们报道了一种合成肽核酸(PNA)与DNA磁性纳米共轭物的替代方法。对130纳米的葡聚糖-磁铁矿颗粒进行化学修饰,以获得含有对细胞环境敏感的可逆键且适合直接引入未修饰寡聚物的多官能化颗粒。由于纳米颗粒的多价性质,当互补的PNA和DNA纳米共轭物混合在一起时,形成的双链结构会导致由沃森-克里克碱基对驱动的纳米颗粒组装。通过光谱学(紫外、傅里叶变换红外)、扫描和透射电子显微镜以及分析有无磁场作用下纳米颗粒共轭物在水溶液中的宏观行为来研究纳米颗粒组装的形成。此外,血清稳定性试验表明,与未共轭的双链相比,PNA/DNA纳米共轭物双链在胎牛血清(FCS)中的酶抗性增强。所描述的纳米系统可扩展到其他双链结构,可能涉及具有生物医学相关性的适体序列,并且在生物技术应用中,为在靶位点获得双链和磁性纳米颗粒的高局部浓度可能非常有用。