Department of Biotechnology and Bioinformatics, Korea University, Sejong 30019, Republic of Korea.
Interdisciplinary Graduate Program for Artificial Intelligence Smart Convergence Technology, Korea University, Sejong 30019, Republic of Korea.
Biosensors (Basel). 2024 Aug 19;14(8):400. doi: 10.3390/bios14080400.
Quantifying the formation and decomposition of amyloid is a crucial issue in the development of new drugs and therapies for treating amyloidosis. The current technologies for grasping amyloid formation and decomposition include fluorescence analysis using thioflavin-T, secondary structure analysis using circular dichroism, and image analysis using atomic force microscopy or transmission electron microscopy. These technologies typically require spectroscopic devices or expensive nanoscale imaging equipment and involve lengthy analysis, which limits the rapid screening of amyloid-degrading drugs. In this study, we introduce a technology for rapidly assessing amyloid decomposition using capillary flow-based paper (CFP). Amyloid solutions exhibit gel-like physical properties due to insoluble denatured polymers, resulting in a shorter flow distance on CFP compared to pure water. Experimental conditions were established to consistently control the flow distance based on a hen-egg-white lysozyme amyloid solution. It was confirmed that as amyloid is decomposed by trypsin, the flow distance increases on the CFP. Our method is highly useful for detecting changes in the gel properties of amyloid solutions within a minute, and we anticipate its use in the rapid, large-scale screening of anti-amyloid agents in the future.
定量研究淀粉样蛋白的形成和分解是开发治疗淀粉样变性新药和疗法的关键问题。目前用于捕捉淀粉样蛋白形成和分解的技术包括使用硫黄素-T 的荧光分析、使用圆二色性的二级结构分析，以及使用原子力显微镜或透射电子显微镜的图像分析。这些技术通常需要分光设备或昂贵的纳米级成像设备，并且涉及冗长的分析，这限制了对淀粉样蛋白降解药物的快速筛选。在这项研究中，我们引入了一种使用基于毛细管流动的纸（CFP）快速评估淀粉样蛋白分解的技术。由于不溶性变性聚合物，淀粉样蛋白溶液表现出凝胶状的物理性质，因此与纯水相比，在 CFP 上的流动距离更短。根据鸡卵清溶菌酶淀粉样蛋白溶液，建立了实验条件以一致地控制基于 CFP 的流动距离。结果证实，随着胰蛋白酶分解淀粉样蛋白，CFP 上的流动距离增加。我们的方法非常适用于在一分钟内检测淀粉样蛋白溶液凝胶性质的变化，我们预计它将在未来用于快速、大规模筛选抗淀粉样蛋白剂。
