Biolayer Interferometry (BLI)

CMI BLI resources and user guide⬇︎

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip.  BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time. Sensors are moved from one well of a microplate to another to change solution between samples (load sample and analyte sample) and buffer for baselines and dissocation.

Biolayer Interferometry Diagram

The CMI has two high sensitivity Sartorius (formerly ForteBio) instruments for measuring BLI of macromolecular and small molecule interactions: the Octet R8e,and the Octet RED384 (now called Octet RH16).  The Octet R8e is more sensitive and has a wider temperature range, whereas the Octet Red384 has a 2x higher throughput. See Octet Instrument Comparison.

 

Applications 

Octet R8e and Red384
  • Kinetic binding: ka, kd
  • Equilibrium binding: KD
  • Macromolecular and small molecule binding

Key  Features

  • Disposable biosensors (regeneration not required)
  • Simultaneous real-time binding on 8 or 16 biosensors
  • 96 or 384 well reagent plate format 

Octet Resources - R8e and Red384(RH16)

BLI Supplies

User-Provided Required supplies:

  • Sartorius Dip and Read sensors*
  • Black 96-well plate to hydrate (soak) sensors**
  • Black 96-well or 384-well plate for samples and reagents**
  • Pipettors for sample handling

Optional provided supplies and equipment:

  • Empty BLI Sensor trays to use as working tray during experiment
  • 8-sensor transfer tool
  • Refrigerated centrifuge with microplate rotor

 

* Sartorius Sensor Selection Guide

** Only Greiner Bio-One brand black polypropylene plates or Sartorius brand black plates are recommended by Sartorius for the Octet.

 

Select Octet Biosensors for Kinetics

TypeSartorius Part Number 
Streptavidin (SA) biosensors 
18-5019   (96/tray)
High Precision Streptavidin (SAX) biosensors
18-5117
Super-Streptavidin (SSA) biosensors (for small molecules)
18-5057
anti-His (HIS1K) biosensors
18-5120
Ni-NTA (NTA) biosensors
18-5101
Anti-Human IgG Fc (AHC) biosensors
18-5060
Anti-Mouse IgG Fc (AMC) biosensors
18-5088
Anti-Flag (DYKDDDDK) Biosensors
18-5187

Octet Black Microplates

TypePart Number 
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 µL
781209 (VWR 82051-318)
Sartorius Octet 384TW microplates, tilted-bottom, 60 µL
18-5166
Sartorius Octet Evaporation Covers
18-5132

For information on access fees, policies and getting started at the CMI, see the CMI Access Page.