Surface plasmon resonance spectroscopy and quartz crystal microbalance study of streptavidin film structure effects on biotinylated DNA assembly and target DNA hybridization
Surface plasmon resonance spectroscopy and quartz crystal microbalance study of streptavidin film structure effects on biotinylated DNA assembly and target DNA hybridization
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DOI：
10.1021/la047997u
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发表时间：
2005-01-04
期刊：
影响因子：
3.9
通讯作者：
Knoll, W
中科院分区：
文献类型：
--
作者：
Su, XD;Wu, YJ;Knoll, W
表面等离子共振 (SPR) 光谱用于研究链霉亲和素修饰金表面上的生物素化 DNA 组装，以实现目标 DNA 杂交。构建链霉亲和素膜涉及两种固定策略，即（1）通过生物素-链霉亲和素连接物理吸附在含生物素硫醇处理的表面上；（2）通过胺偶联共价附着到 11-聚十一烷酸（MUA）处理的表面。了解链霉亲和素薄膜（一种石英晶体）的结构特性。具有能量耗散监测功能的微量天平 (QCM-D) 用于监测链霉亲和素固定程序。同时测量的频率 (Deltaf) 和耗散因子 (AD) 变化以及 SPR 角移 (Deltatheta) 表明，在含生物素的表面上组装的链霉亲和素膜具有高度刚性且结构良好，而通过胺偶联形成的链霉亲和素膜具有高耗散性且结构较差。随后的生物素化DNA（生物素-DNA）组装和靶标杂交结果表明，链霉亲和素膜结构对生物素-DNA结合量有明显影响。在链霉亲和素基质上，不仅探针 DNA 密度而且链霉亲和素膜介导的链方向对杂交效率都有明显影响。特别是，在含生物素的表面上形成的分子有序链霉亲和素薄膜确保了良好有序的DNA组装，与通过胺偶联形成的结构较差的链霉亲和素膜上组装的生物素-DNA相比，这反过来允许更高的目标DNA捕获效率和更高的杂交分析灵敏度。
Surface plasmon resonance (SPR) spectroscopy is employed for the study of biotinylated DNA assembly on streptavidin modified gold surfaces for target DNA hybridization. Two immobilization strategies are involved for constructing streptavidin films, namely, (1) physical adsorption on biotin-containing thiol treated surfaces through biotin-streptavidin links and (2) covalent attachment to 11-mereaptoundecanoic acid (MUA) treated surfaces through amine coupling. To understand the structural properties of the streptavidin films, a quartz crystal. microbalance with energy dissipation monitoring (QCM-D) is used to monitor the streptavidin immobilization procedures. The simultaneously measured frequency (Deltaf) and dissipation factor (AD) changes, together with the SPR angle shifts (Deltatheta), suggest that the streptavidin film assembled on the biotin-containing surface is highly rigid with a well-ordered structure while the streptavidin film formed through amine coupling is highly dissipative and less structured. The subsequent biotinylated DNA (biotin-DNA) assembly and target hybridization results show that the streptavidin film structure has distinct effects on the biotin-DNA binding amount. On the streptavidin matrix, not only the probe DNA density but also the strand orientation mediated by the streptavidin films has distinct effects on hybridization efficiency. Particularly, the molecularly ordered streptavidin films formed on the biotin-containing surfaces ensure a well-ordered DNA assembly, which in turn allows for a higher efficiency in target DNA capture and for a higher sensitivity in the hybridization analysis when compared to the biotin-DNA assembled on the less structured streptavidin films formed through amine coupling.