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.

The CMI has two Sartorius (formerly ForteBio) instruments for measuring BLI: the Octet RED384 (now called Octet RH16) and the BLItz (now called Octet N1).  The Octet RED384 is more sensitive and higher throughput and can be used for macromolecular and small molecule binding experiments.  The BLItz uses a very small sample volume, but is more time consuming and cannot measure molecules smaller than 10 KDa.

Applications and General Features

• Kinetic binding: k_a, k_d
• Equilibrium binding: K_D
• Disposable biosensors (sensor regeneration not required)

Octet RED384 Features

• Simultaneous real-time binding on up to 16 biosensors
• 96 or 384 well reagent plate format
• Small molecule sensitivity

BLItz Feature

• Low sample volume: 4 μl sample