Laboratory of Biomolecular Interactions Studies, Chair of Drug and Cosmetics Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Warsaw, Poland.
Methods Mol Biol. 2022;2393:535-558. doi: 10.1007/978-1-0716-1803-5_28.
The viscoelastic properties of cells are responsible for the adhesion process to different surfaces and for cell motility. Therefore, it is very important to develop specific, label-free biosensors with the use of whole cells to study the effect of various factors on the survival and properties of selected type of normal and pathological cells. The quartz crystal microbalance with dissipation energy monitoring (QCM-D) is a technique which enables to track these changes in cells during real-time experiments. One of the applied procedures of the evaluation of the cells' viscoelastic changes is based on the investigations of interactions between specific, different glycans, present on the surface of the primary tumor and its metastases with specific lectins. Two procedures have been developed to detect the differences in the cellular glycosylation profile using cell-based sensors (adherent cells cultured on sensors) and suspension cell-based sensors (adherent cells mechanically detached and inserted into the QCM-D chamber with a sensor). Furthermore, in this work some cell-based sensor regeneration protocols have been described and a lectin-ELISA assay with a fluorescently labeled lectin, thus enabling a qualitative and quantitative tracking of each step of the lectin-glycan binding and unbinding process performed on whole cells.
细胞的黏弹性特性是细胞黏附到不同表面和细胞运动的原因。因此，开发使用全细胞的特定、无标记的生物传感器来研究各种因素对选定类型的正常和病理细胞的存活和特性的影响非常重要。石英晶体微天平耗散能量监测（QCM-D）是一种能够在实时实验中跟踪细胞这些变化的技术。评估细胞黏弹性变化的应用程序之一是基于研究原发性肿瘤及其转移灶表面特定、不同糖链与特定凝集素之间的相互作用。已经开发了两种程序来使用基于细胞的传感器（在传感器上培养的贴壁细胞）和悬浮细胞基于传感器（机械分离的贴壁细胞并插入 QCM-D 室中的传感器）来检测细胞糖基化谱的差异。此外，在这项工作中，描述了一些基于细胞的传感器再生方案，并进行了带有荧光标记凝集素的凝集素-ELISA 测定，从而能够定性和定量跟踪在整个细胞上进行的凝集素-糖结合和解离过程的每一步。
