Kwon Sung Hong, Hong Bong Jin, Park Hye Young, Knoll Wolfgang, Park Joon Won
Center for Integrated Molecular Systems, Department of Chemistry, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea.
J Colloid Interface Sci. 2007 Apr 15;308(2):325-31. doi: 10.1016/j.jcis.2007.01.041. Epub 2007 Jan 23.
Since we observed that dendron-assembled surface provided high single nucleotide polymorphism discrimination efficiency for DNA microarrays, and that the binding yield for streptavidin increased when biotin was immobilized on top of it, the nanoscale-controlled surface is examined for surface plasmon field-enhanced fluorescence spectroscopy (or SPFS). Firstly, a silica film was coated onto a gold substrate using the sol-gel technique, followed by the covalent immobilization of a layer of second-generation dendrons with a DNA catcher strand at their apex. The thickness of the inorganic interlayer (d=33 nm) was effectively suppressing fluorescence quenching. Thus, the kinetics and affinity characteristics of DNA hybridization could be investigated very sensitively by SPFS. The kinetic rate constants found for DNA hybridization on the dendron-modified surface were larger than those reported for a streptavidin-modified surface by one order of magnitude, except for dissociation rate constant for a single mismatched case. In addition, we observed that the DNA on the cone-shaped linker maintained its capability to capture DNA target strands even after extended storage at ambient conditions.
由于我们观察到树枝状分子组装表面为DNA微阵列提供了高单核苷酸多态性鉴别效率,并且当生物素固定在其顶部时链霉亲和素的结合产率增加,因此对这种纳米级可控表面进行了表面等离子体场增强荧光光谱(或SPFS)研究。首先,使用溶胶-凝胶技术在金基底上涂覆一层二氧化硅膜,然后在其顶端通过共价固定一层带有DNA捕获链的第二代树枝状分子。无机中间层的厚度(d = 33 nm)有效抑制了荧光猝灭。因此,通过SPFS可以非常灵敏地研究DNA杂交的动力学和亲和特性。除了单碱基错配情况下的解离速率常数外,在树枝状分子修饰表面上发现的DNA杂交动力学速率常数比链霉亲和素修饰表面报道的速率常数大一个数量级。此外,我们观察到即使在环境条件下长时间储存后,锥形连接体上的DNA仍保持捕获DNA靶链的能力。