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Development of Surface Plasmon Resonance-type Quartz Crystal Microbalance and Detection of Interactions of Viomolecules

表面等离激元共振型石英晶体微天平的研制及分子相互作用的检测

基本信息

批准号：
15201025
负责人：
OKAHATA Yoshio
金额：
$ 32.36万
依托单位：
Tokyo Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

By using a 27-MHz piezoelectric quartz oscillator connected with a vector network analyzer, we obtained resonance frequency decreases (-ΔF_<water>) and energy dissipation increases (ΔD_<water>) during binding of biotinylated BSA (bovine serum albumin), biotinylated ssDNA, biotinylated dsDNA, and biotinylated pullulan to a NeutrAvidin-immobilized 27-MHz QCM (quartz crystal microbalance) plate in aqueous solution, as well as in the wet air phase (97% humidity, -ΔF_<wet> and ΔD_<wet>) and in the dry air phase (-ΔF_<air> and ΔD_<air>). The -ΔF_<air> value indicates real mass changes due to the binding of these biomolecules. Values for polystyrene were also obtained as elastic and hydrophobic membranes. In terms of results, (-ΔF_<wet>)/(-ΔF_<air>) values indicated the binding water ratio per unit biomolecular mass were in the order of pullulan (2.1- 2.2) > DNAs = proteins (1.4- 1.6) > polystyrene (1.0). The (-ΔF_<water>)/(-ΔF_<air>) values indicated the hydrodynamic water (binding and vibrating water) ratio per unit biomolecular mass were in the order of dsDNA (6.5)> ssDNA = pullulan (3.5- 4.4) > proteins (2.4- 2.5) > polystyrene (1.0). The aforementioned values suggest that the energy dissipation of proteins was mainly caused by hydration and that proteins themselves are elastic molecules without energy dissipation in aqueous solutions. On the contrary, plots in cases of DNAs and pullulans were relatively deviant toward the large hydration and energy dissipation from the linearity observed with proteins, meaning that the large energy dissipation occurs because of viscoelastic properties of linear DNAs and pullulans in the water phase, in addition to energy dissipation due to the hydration of molecules.

利用27-MHz压电石英振荡器与矢量网络分析仪连接，我们获得了生物素化BSA（牛血清白蛋白）、生物素化ssDNA、生物素化dsDNA和生物素化pululan与固定化neutravidin的27-MHz QCM（石英晶体微天平）板在水溶液中以及湿气相（97%湿度，97%湿度）结合时的共振频率降低（-ΔF_<water>）和能量耗散增加（ΔD_<water>）。-ΔF_<wet>和ΔD_<wet>)和在干气相（-ΔF_<air>和ΔD_<air>）。-ΔF_<air>值表示由于这些生物分子的结合而导致的实际质量变化。聚苯乙烯作为弹性膜和疏水膜的性能也得到了数值。结果表明，（-ΔF_<wet>）/（-ΔF_<air>）表示单位生物分子质量的结合水比依次为普鲁兰(2.1- 2.2)> dna =蛋白质(1.4- 1.6)>聚苯乙烯（1.0）。（-ΔF_<water>）/（-ΔF_<air>）表示单位生物分子质量的水动力水（结合水和振动水）比依次为dsDNA (6.5)> ssDNA =普鲁兰(3.5 ~ 4.4)>蛋白(2.4 ~ 2.5)>聚苯乙烯（1.0）。上述数值说明蛋白质的能量耗散主要是由水合作用引起的，蛋白质本身是弹性分子，在水溶液中没有能量耗散。相反，dna和普鲁兰的图相对偏离了与蛋白质的线性关系所观察到的大水化和能量耗散，这意味着除了分子水化引起的能量耗散外，还由于线性dna和普鲁兰在水相中的粘弹性而产生了大的能量耗散。