Resoruce Overview
Low-affinity interactions have traditionally been assessed using equilibrium and endpoint assays due to a lack of assay resolution. The use of equilibrium-based assays and steady-state analysis has several limitations in accuracy and precision due to the mathematical model used to fit the data, especially when assessing diverse molecules. Here, we demonstrate how the enhanced data acquisition rate on the Octet® R8e can be used to generate more accurate data for low-affinity interactions across a range of temperatures relative to steady-state analysis.
Resoruce Details:
- Document Type: Application Note
- Page Count: 12
- Read Time: 22 minutes
Key Takeaways
- Enhanced Accuracy: Kinetic analysis provides precise interaction dynamics using association and dissociation rates, improving accuracy compared to steady-state analysis.
- Detailed Binding Characteristics: It uncovers binding differences that steady-state models might miss, offering deeper insights into CD16b NA1 and NA2 variants.
- Real-Time Monitoring: BLI technology enables real-time interaction monitoring, crucial for understanding dynamic interaction behavior across temperatures.
- Thermodynamic Insights: Analyzing interactions at various temperatures reveals thermodynamic properties and interaction dynamics, aiding drug design and formulation.
- Robustness and Reliability: The kinetic model consistently delivers accurate affinity values across temperatures, demonstrating superiority over steady-state analysis.