Huang Chih-Ching, Chiu Sheng-Hsien, Huang Yu-Fen, Chang Huan-Tsung
Department of Chemistry, National Taiwan University, 1, Section 4, Roosevelt Road, Taipei 106, Taiwan.
Anal Chem. 2007 Jul 1;79(13):4798-804. doi: 10.1021/ac0707075. Epub 2007 May 26.
An aptamer modified gold nanoparticles (Apt-AuNPs) based molecular light switching sensor has been demonstrated for the analysis of breast cancer markers (platelet-derived growth factors (PDGFs) and their receptors) in homogeneous solutions. The PDGF binding aptamer has a unique structure with triple-helix conformation that allows N,N-dimethyl-2,7-diazapyrenium dication (DMDAP) and PDGF bindings. The fluorescence of DMDAP is almost completely quenched by Apt-AuNPs when it intercalates with the aptamers. Owing to high magnitudes of increases (up to 40-fold) in the turn-on fluorescence signals of DMDAP/Apt-AuNP upon PDGFs binding, the approach is highly sensitive for the detection of PDGFs. The DMDAP/Apt-AuNP probe specifically and sensitively detected PDGFs under optimal concentrations of salts and DMDAP. We also demonstrated that the Apt-AuNPs are effective selectors for enrichment of PDGF-AA from large-volume samples. The approach allows detection of PDGF-AA at a concentration down to 8 pM, showing better sensitivity than other signal aptamers. By conducting a competitive assay, we demonstrated the determination of PDGF receptor-alpha with LOD of 0.25 nM when using the DMDAP/Apt-AuNP as a probe.
一种基于适配体修饰金纳米颗粒(Apt-AuNPs)的分子光开关传感器已被用于在均相溶液中分析乳腺癌标志物(血小板衍生生长因子(PDGFs)及其受体)。PDGF结合适配体具有独特的三螺旋构象结构,允许N,N-二甲基-2,7-二氮杂芘二价阳离子(DMDAP)与PDGF结合。当DMDAP与适配体插入时,其荧光几乎被Apt-AuNPs完全淬灭。由于PDGFs结合后DMDAP/Apt-AuNP的开启荧光信号有大幅增加(高达40倍),该方法对PDGFs的检测高度灵敏。DMDAP/Apt-AuNP探针在最佳盐浓度和DMDAP浓度下能特异性且灵敏地检测PDGFs。我们还证明了Apt-AuNPs是从大量样品中富集PDGF-AA的有效选择剂。该方法能够检测低至8 pM浓度的PDGF-AA,显示出比其他信号适配体更好的灵敏度。通过进行竞争性测定,我们证明了使用DMDAP/Apt-AuNP作为探针时,PDGF受体-α的检测限为0.25 nM。