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Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
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A novel label-free cell-based assay technology using biolayer interferometry.

D Verzijl1, T Riedl1, P W H I Parren2

  • 1Genmab, Yalelaan 60, Utrecht, 3584 CM, The Netherlands.

Biosensors & Bioelectronics
|September 3, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces cell-based biolayer interferometry (cBLI) for real-time, label-free monitoring of signal transduction in living cells. The technique tracks dynamic mass redistribution, enabling efficient compound screening and characterization.

Keywords:
Biolayer interferometryBiosensorCell-basedDynamic mass redistributionForteBio Octet HTXLabel-free

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Biophysics

Background:

  • Biolayer interferometry (BLI) is a label-free optical technique for studying biomolecular interactions.
  • Real-time monitoring of cellular signal transduction typically requires labeled probes.

Purpose of the Study:

  • To develop and validate a cell-based BLI (cBLI) application for label-free, real-time monitoring of signal transduction in living cells.
  • To demonstrate cBLI's utility as a cellular assay for compound screening and characterization.

Main Methods:

  • Human A431 epidermoid carcinoma cells were cultured on collagen-coated biosensors.
  • Cells were stimulated with various agonists targeting epidermal growth factor receptor (EGFR), β2-adrenoceptor, and hepatocyte growth factor receptor (HGFR/c-MET).
  • cBLI signals were recorded in real-time, and changes were correlated with actin cytoskeleton rearrangement (dynamic mass redistribution - DMR).

Main Results:

  • Distinct cBLI signal patterns were observed upon stimulation of different receptors (EGFR, β2-adrenoceptor, HGFR/c-MET).
  • The observed cBLI signal changes were mediated by dynamic mass redistribution (DMR) involving actin cytoskeleton rearrangement.
  • The assay successfully differentiated between ligand-binding and non-binding anti-EGFR antibodies.

Conclusions:

  • Cell-based BLI (cBLI) provides a novel, label-free method for real-time monitoring of signal transduction in living cells.
  • cBLI effectively detects receptor-mediated cellular responses, including dynamic mass redistribution.
  • This technique is suitable for label-free cellular assays in compound screening and characterization.