Issue link: https://resources.nanotempertech.com/i/1533610
6 A P P L I C A T I O N N O T E Hans-Jörg emphasizes that Spectral Shi technology provides a critical advantage when working with high concentrations of compounds, which typically interfere with traditional assay systems. "In methods like SPR, if we had to increase compound concentration above 50 μM, we would start to see undesired artifacts," Hans-Jörg says. In contrast, Spectral Shi allows for accurate readings, even at much higher compound concentrations, maintaining the integrity of the data. These characteristics give Spectral Shi technology the ability to evaluate the cooperativity of proximity inducers at a broader range of concentrations, which is critical for monitoring cooperativity through lower affinity binding proteins. Hans-Jörg also highlights the importance of sensitivity in proximity inducer screening and characterization, noting that Dianthus' Spectral Shi technology excels in this area. "Sensitivity is also key. In cases where cooperativity 2 is measured through the stronger binding protein, we need the ability to work with protein concentrations sufficiently under the sometimes low ternary K d s," Hans-Jörg states. This dual characteristic — sensitivity to low protein concentrations while also discerning weak interactions — enables the determination of cooperativity through the weaker and/or the stronger binding protein, which is a great way to validate the obtained data within the same system. Moreover, Dianthus addresses the need for an in-solution method, which allows for more precise control of protein concentrations compared to immobilization-based techniques. "When immobilizing a protein on a surface, it's difficult to know the exact concentration," Hans-Jörg points out, "but with solution- based assays, we can control the exact protein concentrations, which is essential for accurately assessing cooperativity." Lastly, Dianthus supports medium-throughput screening 3 , which is ideal for hit characterization following high-throughput screening (HTS). "A er screening hundreds of thousands of compounds, the resulting hit list can contain hundreds or even thousands of hits," Hans- Jörg notes, "Dianthus allowed us to handle that volume effectively without losing precision, making it a valuable tool for hit validation and further characterization." Furthermore, the system's low sample volume requirements make it a cost-effective solution, as large volumes of counter-protein can be expensive. "Having low sample volume while still achieving accurate results is crucial," Hans-Jörg adds. In summar y, Dianthus with Spectral Shift technology provides a unique solution to the challenges of screening and characterizating proximity inducers. By offering a high sensitivity, low sample volume, in-solution method with precise protein concentration control, 2 The absolute (not the apparent) cooperativity α is the same, regardless of the direction of monitoring of binding. U S E R C A S E S T U D Y