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Protein-protein Interaction Study

Protein Core

Introduction

The VIB Protein Core offers a study of your protein of interests interactions with other proteins using our Bio-Layer Interferometry instrument. We can determine the kinetic parameters of your protein using this technique. It is often used to determine dissociation constants for nanobodies or antibodies. It gives a good idea of the binding rates and strength of the binding in order to validate your protein.

OctetRed96 (Source: Sartorius - Moldev - Fortébio)

Bio-Layer Interferometry

Bio-Layer Interferometry (BLI) is a label-free technology for measuring biomolecular interactions. It is an optical analytical technique that analyzes the interference pattern of white light reflected from 2 surfaces:

  • A layer of immobilized protein on the biosensor tip
  • An internal reference layer

BLI interference pattern (Source:https://www.fortebio.com/applications/bli-technology)

If now a ligand is coupled to the biosensor and its analyte binds the ligand, this produces an increase in optical thickness at the biosensor tip, which results in a wavelength shift (Δλ). This is a direct measure of the change in thickness of the biological layer.

These interactions are measured in real-time with high precision and accuracy, providing the ability to monitor:

  • Binding specificity
  • Rates of association and dissociation
  • Concentration

Only molecules binding to or dissociating from the biosensor can shift the interference pattern and generate a response. Unbound molecules, changes in refractive index (RI) of the medium or changes in flow rate do not affect the interference pattern. Which makes it suitable to measure even in crude samples for protein-protein binding, quantitation, affinity and kinetics.

Binding profile (Source: Sartorius - Moldev - FortéBio)