Binding kinetics make up the foundation of pharmacological theory in the modern era. Binding kinetics is a term that is concerned with the speed at which a compound binds to a target and how quickly it dissociates from it.
BLI (bio-layer interferometry) is an optical biosensing technology used in analyzing biomolecular interactions without requiring fluorescent labeling. BLI is one of the few widely available biosensing technologies that are label-free. BLI measures macromolecular interactions by analyzing the patterns of interference from white light reflected from a biosensor tip surface. This article discusses the benefits of BLI for measuring binding kinetics.
What Parameters of Binding Kinetics can be Determined Using BLI?
BLI allows for real-time determination of association rate (kon) and dissociation rate (koff) in molecular interaction and as such the binding kinetics. These rates provide the opportunity to understand the overall affinity (KD) of the interaction to be calculated. This value can also be ascertained directly from the formation of concentration-dependent molecular complexes.
Benefits of BLI
Simplicity
One key benefit of using BLI for binding kinetics measurements is that Kon, Koff, and KD can be measured in one rapid assay. Detection takes place in real-time without marking biological molecules. This makes the measurement quicker and simpler than processes that require multiple assays.
Wide Ranges
Using BLI for binding kinetics allows for a wide range of affinity values. BLI can be used for many investigations such as nanobodies, membrane proteins, antibodies, AAV, small molecules, nucleic acids, and more.
Compatible with a Range of Samples
BLI is compatible with both crude and purified samples. This means it can be used in a range of investigations and there is no need to swap between systems for samples.
No Labels Required
Label-free techniques can monitor responses in real-time, meaning kinetic information regarding dissociation and association is accessible. They can also monitor bio-molecular interactions with native binding partners without any impediment from modified analytes and therefore more accurate biochemical data is acquired.
BLI for Binding Kinetics
Gator Bio software provides a range of models for data analysis. Reference subtraction, processing, and data fitting can be performed quickly. Biosensors from Gator Bio combine a 1mm diameter glass rod with patented optical layers and surface chemistry specifically built at the distal end of the biosensor. To find out more about how this technology can be used in binding kinetics applications, contact the team at Gator Bio today.