Buell Alexander K, Dobson Christopher M, Welland Mark E
Nanoscience Centre, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2012;849:101-19. doi: 10.1007/978-1-61779-551-0_8.
Kinetic measurements of amyloid growth provide insight into the free energy landscape of this supramolecular process and are crucial in the search for potent inhibitors of the main disorders with which it is associated, including Alzheimer's and Parkinson's diseases and Type II diabetes. In recent years, a new class of surface-bound biosensor assays, e.g., those based on surface plasmon resonance (SPR) and the quartz crystal microbalance (QCM) have been established as extremely valuable tools for kinetic measurements of amyloid formation. Here we describe detailed protocols of how QCM techniques can be used to monitor the elongation of amyloid fibrils in real time and to study the influence of external factors on the kinetics of amyloid growth with unprecedented accuracy.
淀粉样蛋白生长的动力学测量有助于深入了解这一超分子过程的自由能景观,对于寻找与之相关的主要疾病(包括阿尔茨海默病、帕金森病和II型糖尿病)的有效抑制剂至关重要。近年来,一类新型的表面结合生物传感器检测方法,例如基于表面等离子体共振(SPR)和石英晶体微天平(QCM)的检测方法,已成为淀粉样蛋白形成动力学测量的极有价值的工具。在此,我们描述了详细的实验方案,介绍如何使用QCM技术实时监测淀粉样纤维的伸长,并以前所未有的精度研究外部因素对淀粉样蛋白生长动力学的影响。