Method
Label-free optical detection
Data
Real-time kinetics
Sensitivity
Sub-nanomolar detection
1
Ligand Immobilization
Designed proteins immobilized on biosensor tip using Twin-Strep-Tag
2
Analyte Binding
Target protein flows over sensor surface
3
Signal Detection
Binding causes detectable light interference shift
4
Kinetic Analysis
Association and dissociation rates calculated
Key Parameters
BLI provides three critical binding parameters: k_on (association), k_off (dissociation), and K_D (equilibrium constant).
- k_on - Association rate constant
- k_off - Dissociation rate constant
- K_D - Equilibrium dissociation constant (k_off/k_on)
How we use BLI at Adaptyv
We use the Gator Bio Pro for BLI measurements. Our typical workflow involves:- Protein immobilization: User-designed proteins are expressed with a linker and Twin-Strep-Tag
- Target protein binding: Biosensor is moved into solutions of the target protein at different concentrations
- Kinetic analysis: Real-time binding measurements to determine association and dissociation kinetics