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BIO-LAYER INTERFEROMETRY (BLI)

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Bio-layer interferometry (BLI)

Bio-Layer Interferometry (BLI), a label-free technology that measures molecular interactions in real time for the purpose of detection, quantitation and kinetic analysis. BLI experiments are used to determine the kinetics and affinity of molecular interactions.

BLI is an optical analytical technique that measures interferfence patterns between waves of light. White light is directed down the fiber-optic biosensor towards two interfaces separated by a thin layer at the tip of the fiber: a biocompatible layer on the surface of the tip, and an internal reference layer. Light reflects from each of the two layers, and the reflected beams interfere constructively or destructively at different wavelengths in the spectrum. This interference pattern is detected at the CCD array detector.

In a BLI experiment, one molecule (the Load Sample) is immobilized to a Dip and Read Biosensor and binding of a second molecule (the Analyte Sample) is then measured. This binding forms a molecular layer that increases in thickness as more molecules bind to the surface. As the thickness at the tip increases, the effective distance between the two reflective layers increases, creating a shift in the interference pattern of the reflected light. The spectral pattern of the reflected light therefore changes as a function of the thickness of the molecular layer, i.e., the number of molecules bound to the biosensor surface. Response is measured as a nm shift in the interference pattern and is proportional to the number of molecules bound to the surface of the biosensor. This spectral shift is monitored at the detector, and reported on a sensorgram as a change in wavelength (nm shift). Monitoring the interference pattern in real time provides kinetic data on molecular interactions.

Octet BLI System      Octet biomolecular binding kinetics application

Instrument Overview

The IBV has an Octet R8 instrument from Sartorius, which measures binding kinetics and equilibrium by BLI using ForteBio dip and read biosensors and black microplates. The Octet R8 system can run up to eight samples in parallel.

octet r brochure en l 4047

Required Supplies

  • ForteBio Biosensors

Biosensors used with the Octet system are single-use. The tip of each fiber-optic biosensor is coated with a biocompatible matrix that minimizes non-specific binding to the surface. This matrix is coated with one of a wide selection of biosensor chemistries available for highly specific binding of target molecules in a sample. Regeneration of the biosensors is possible in many cases.

Biosensor selection compatibility guide

  • Black microplates

At least two per experiment (one for soaking sensors and at least one for samples and reagents). Only Greiner Bio-One brand, black microplates or ForteBio plates are recommended.

Greiner Bio-One 96-well black flat-bottom PP, 200 µL 655209 (VWR 82050-784)

Greiner Bio-One 384-well black flat-bottom PP, 80-120 µL 781209 (VWR 82051-318)

ForteBio 384-well black tilted-bottom PP, 60 µL 18-5080

  • An empty biosensor tray to use as a working tray.

 

Manuals

  • BLI getting started guide

  • Octet BLI Discovery and OctetAnalysis Studio Software

Octet Software

 

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