Grundmann Manuel
Section Cellular, Molecular and Pharmacobiology, Institute for Pharmaceutical Biology, University of Bonn, Bonn, Germany.
Curr Protoc Pharmacol. 2017 Jun 22;77:9.24.1-9.24.21. doi: 10.1002/cpph.24.
Label-free biosensors are increasingly employed in drug discovery. Cell-based biosensors provide valuable insights into the biological consequences of exposing cells and tissues to chemical agents and the underlying molecular mechanisms associated with these effects. Optical biosensors based on the detection of dynamic mass redistribution (DMR) and impedance biosensors using cellular dielectric spectroscopy (CDS) capture changes of the cytoskeleton of living cells in real time. Because signal transduction correlates with changes in cell morphology, DMR and CDS biosensors are exquisitely suited for recording integrated cell responses in an unbiased, yet pathway-specific manner without the use of labels that may interfere with cell function. Described in this unit are several experimental approaches utilizing optical label-free system capturing dynamic mass redistribution (DMR) in living cells (Epic System) and an impedance-based CDS technology (CellKey). In addition, potential pitfalls associated with these assays and alternative approaches for overcoming such technical challenges are discussed. © 2017 by John Wiley & Sons, Inc.
无标记生物传感器在药物研发中的应用日益广泛。基于细胞的生物传感器能为探究细胞和组织接触化学试剂后的生物学后果以及这些效应背后的分子机制提供有价值的见解。基于动态质量再分布（DMR）检测的光学生物传感器和使用细胞介电谱（CDS）的阻抗生物传感器可实时捕捉活细胞细胞骨架的变化。由于信号转导与细胞形态变化相关，DMR和CDS生物传感器非常适合以无偏倚但具有通路特异性的方式记录整合细胞反应，且无需使用可能干扰细胞功能的标记物。本单元介绍了几种利用光学生物无标记系统捕捉活细胞动态质量再分布（DMR）（Epic系统）和基于阻抗的CDS技术（CellKey）的实验方法。此外，还讨论了与这些检测相关的潜在陷阱以及克服此类技术挑战的替代方法。© 2017约翰威立国际出版公司
