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

• 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
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15201025/
• 关键词: Quartz crystal microbalance; Network analyzer; Energy dissipation factor; Conformation of Proteins; p53 Protein; タンパク質のコンフォメーション変化; 表面プラズモン共鳴; インピーダンスアナライザー; 分子認識; Quartz crystal microbalance; Network analyzer; Energy dissipation factor; Conformation of Proteins; p53 Protein; タンパク質のコンフォメーション変化; 表面プラズモン共鳴; インピーダンスアナライザー; 分子認識

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.

通过使用与矢量网络分析仪相关的27-MHz压电石英振荡器，我们在生物素化BSA（牛血清白蛋白）的生物素化BSA结合期间获得了共振频率下降（-ΔF_<Water>）和能量耗散的增加（ΔD_D_<water>）中性蛋白含量的27-MHz QCM（石英晶体微量平衡）在水溶液中以及湿空气相（97％湿度，-ΔF_<wew>和ΔD_<wew>）和干燥空气相（-ΔF_F_F_F_F_F_F_<iir>和ΔD_D_<ir>）。 -ΔF_<iar>值表示由于这些生物分子的结合而导致的实际质量变化。聚苯乙烯的值也获得为弹性和疏水膜。在结果方面，（-ΔF_<weg>）/（ - ΔF_<air>）值表明每单位生物分子质量的结合水比为pullulan（2.1- 2.2）> dnas = dnas =蛋白质（1.4-1.6）>聚苯乙烯（1.0）。 （-ΔF_<water>）/（ - ΔF_<iar>）值表示每单位生物分子质量的流体动力水（结合和振动水）的比例为dsDNA（6.5）> ssDNA = pluplulan（3.5-4.4）> proultein（3.5- 4.4）>蛋白质（2.4-2.5）> 2.4-2.5）>> Polystyrene（2.4-2.5）> Polystyrene（1.0）。理性值表明蛋白质的能量耗散主要是由水合引起的，并且蛋白质本身是水溶液中没有能量耗散的弹性分子。在形成鲜明的情况下，在DNA和Pullulans的情况下，与用蛋白质观察到的线性的较大的水合和能量耗散相对偏见，这意味着由于分子水化而导致的能量耗散，由于水相中的线性DNA和Pullulans的粘弹性耗散而发生。