A novel label-free cell-based assay technology using biolayer interferometry

作者：Verzijl D; Riedl T; Parren P W H I; Gerritsen A F^*

来源：Biosensors and Bioelectronics, 2017, 87: 388-395.

DOI：10.1016/j.bios.2016.08.095

• Biolayer interferometry
• Cell-based
• Label-free
• Dynamic mass redistribution
• Biosensor
• ForteBio Octet HTX

Biolayer interferometry (BLI) is a well-established optical label-free technique to study biomolecular interactions. Here we describe for the first time a cell-based BLI (cBLI) application that allows label-free real-time monitoring of signal transduction in living cells. Human A431 epidermoid carcinoma cells were captured onto collagen-coated biosensors and serum-starved, followed by exposure to agonistic compounds targeting various receptors, while recording the cBLI signal. Stimulation of the epidermal growth factor receptor (EGFR) with EGF, the beta(2)-adrenoceptor with dopamine, or the hepatocyte growth factor receptor (HGFR/c-MET) with an agonistic antibody resulted in distinct cBLI signal patterns. We show that the mechanism underlying the observed changes in cBLI signal is mediated by rearrangement of the actin cytoskeleton, a process referred to as dynamic mass redistribution (DMR). A panel of ligand-binding blocking and non-blocking anti-EGFR antibodies was used to demonstrate that this novel BLI application can be efficiently used as a label-free cellular assay for compound screening and characterization.

• 出版日期2017-1-15