Biolayer Interferometry (BLI)

Applications

• Kinetic binding: k_a, k_d
• Equilibrium binding: K_D
• Macromolecular and small molecule binding (Octet only)

Octet RED384 Key Features

• Simultaneous real-time binding on up to 16 biosensors
• Disposable biosensors (sensor regeneration not required)
• 96 or 384 well reagent plate format
• Small molecule sensitivity

BLItz Key Features

• Low sample volume: 4 μl sample
• Disposable biosensors (sensor regeneration not required)

 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 from samples (load sample and analyte sample) to buffer for baselines and dissocation.

Figure adapted from ForteBio. 

There are two major phases in a BLI experiment and each will need to be optimized for you experiment. Regeneration is an optional third phase that may be used when a single set of sensors is reused for multiple measurements.

Immobilization

• the Load molecule is attached irreversibly or reversibly to the sensor chip surface

Interaction analysis

• In the association phase, the Analyte is injected over the sensor surface
• In the dissociation phase, the Analyte is washed off the surface

Regeneration

• the surface is regenerated by removing remaining bound analyte or by removing ligand (which will also remove remaining analyte)

Users are responsible for storage of all raw and processed data collected at the CMI.

• Users should have a plan to copy or transfer all raw and process data to their own local or cloud storage system.
• While the CMI allows temporary local storage of CMI User data on the instrument computer, we make no guarantees on the security or long-term availability of any data at the CMI.
• For most (but not all) CMI technologies, the raw data files and recommended readable exports are relatively small and can be readily transferred electronically. 
• See specific instruments for exceptions and for details about the software, data file types and recommended data exports. 

Data Sharing:

• Currently, a Generalist Repository is the recommended data repository for most CMI data types, as stable specialist data repositories have not been established.

CMI Research Resource Identifier: RRID:SCR_018270

• If you use any CMI resources, please acknowledge the "Center for Macromolecular Interactions at Harvard Medical School, RRID:SCR_018270" in publications of your work.