title: A novel label-free cell-based assay technology using biolayer interferometry.
abstract: 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 β2-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.
authors: Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands.
authors: D
authors: D
authors: Verzijl
authors: Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands.
authors: T
authors: T
authors: Riedl
authors: Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands; Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Albinusdreef 2, Leiden, 2333 ZA, The Netherlands.
authors: P W H I
authors: PWHI
authors: Parren
authors: Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands. Electronic address: A.Gerritsen@genmab.com.
authors: A F
authors: AF
authors: Gerritsen
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D018914:Cells, Immobilized / Q000166:cytology / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D066246:ErbB Receptors / Q000819:agonists / Q000378:metabolism; D006801:Humans; D007368:Interferometry / Q000379:methods*; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism
chemical_list: D018343:Receptors, Adrenergic, beta-2; C512478:EGFR protein, human; D066246:ErbB Receptors; D019859:Proto-Oncogene Proteins c-met
keywords: Biolayer interferometry; Biosensor; Cell-based; Dynamic mass redistribution; ForteBio Octet HTX; Label-free