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

Autor: Verzijl D; Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands., Riedl T; Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands., Parren PWHI; 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., Gerritsen AF; Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands. Electronic address: A.Gerritsen@genmab.com.
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2017 Jan 15; Vol. 87, pp. 388-395. Date of Electronic Publication: 2016 Aug 28.
DOI: 10.1016/j.bios.2016.08.095
Abstrakt: 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 β 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.
(Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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