AbstractNanoTemper Technologies presents a novel technological advancement to quantify molecular interactions in solution, known as ligand-induced spectral shift technology. When a target is labelled with a fluorophore, it generates a particular emission spectrum. If a ligand binds to this labelled target, the fluorophore’s chemical environment is changed, causing a shift in fluorescence spectra. Ratiometric measurement of two emission wavelengths is used to derive the affinity constant (Kd) which is automatically determined at the end of each run.
Ligand-induced spectral shift detection enables characterisation of in solution interactions for a wide range of biomolecules, even for challenging samples such as membrane proteins, intrinsically disordered proteins, and cell lysates.
Since the binding partners are in solution, there is no lost activity due to immobilization, and evaluation is size independent. Measurements can be performed in any buffer, including detergents, using low sample volumes and concentrations. The spectral shift technology also facilitates the evaluation of competition assays and ternary binding events.
With this novel technology, there is no need for multiple rounds of assay development as interactions are detected even in suboptimal buffer conditions and measurements only take a few seconds to generate data.
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