High-throughput investigation of macromolecular interactions for drug development using spectral shift technology

This review focuses on spectral shift analysis as a tool to study macromolecular interactions and describes its current place among the available biophysical methods. NanoTemper's Dianthus platform facilitates a plate-based, microfluidics-free, mass-independent, and immobilisation-free high-throughput screening platform for protein-ligand, protein-protein, and protein-nucleic acid interactions, as well as ternary complexes, for example in proteolysis targeting chimera (PROTAC) design. In addition to spectral shift, the Dianthus offers an orthogonal method, temperature-related intensity change (TRIC). Both methods are presented alongside fluorescent labelling techniques. Specific examples with practical tips for spectral shift methods for diverse binding partners are provided. Finally, current and future applications of spectral shift methods in the drug discovery process are discussed in the context of high-throughput screening, fragment-based drug discovery, and hit-to-lead optimisation.