Mateo-Martí E, Briones C, Pradier C M, Martín-Gago J A
Centro de Astrobiología (CSIC-INTA), Madrid, Spain.
Biosens Bioelectron. 2007 Apr 15;22(9-10):1926-32. doi: 10.1016/j.bios.2006.08.012. Epub 2006 Sep 25.
We present a DNA biosensor based on self-assembled monolayers (SAMs) of thiol-derivatized peptide nucleic acid (PNA) molecules adsorbed on gold surfaces. Previous works have shown that PNA molecules at an optimal concentration can be self-assembled with their molecular axes normal to the surface. In such structural configuration BioSAMs of PNAs maintain their capability for recognizing complementary DNA. We describe the combined use of PM-RAIRS and synchrotron radiation XPS for the detection and spectroscopic characterization of PNA-DNA hybridization process on gold surfaces. RAIRS and XPS are powerful techniques for surface characterization and molecular detection, which do not require a fluorescence labeling of the target. We present a characterization of the spectroscopic IR and XPS features, some of them associated to the phosphate groups of the DNA backbone, as an unambiguous signature of the PNA-DNA heteroduplex formation. The N(1s) XPS core level peak after DNA hybridization is decomposed in curves components, and every component assigned to different chemical species. Therefore, the results obtained by means of two complementary structural characterization techniques encourage the use of PNA-based biosensors for the detection of DNA molecules on natural samples.
我们展示了一种基于吸附在金表面的硫醇衍生化肽核酸(PNA)分子自组装单层(SAMs)的DNA生物传感器。先前的研究表明,最佳浓度的PNA分子可以以其分子轴垂直于表面的方式进行自组装。在这种结构配置下,PNA的生物自组装单层保持其识别互补DNA的能力。我们描述了结合使用偏振调制反射红外光谱(PM-RAIRS)和同步辐射X射线光电子能谱(XPS)来检测和光谱表征金表面上PNA-DNA杂交过程。RAIRS和XPS是用于表面表征和分子检测的强大技术,不需要对目标进行荧光标记。我们展示了光谱红外和XPS特征的表征,其中一些与DNA主链的磷酸基团相关,作为PNA-DNA异源双链形成的明确标志。DNA杂交后的N(1s) XPS核心能级峰被分解为曲线成分,每个成分对应不同的化学物种。因此,通过两种互补的结构表征技术获得的结果鼓励使用基于PNA的生物传感器来检测天然样品中的DNA分子。