Molecules that bind to targets with high specificity and affinity are used as therapeutic and diagnostic agents in medicine and research. Protein and non-protein ligands are used for introduction of mutations within the binding region to increase the binding affinity. Phage-display technology is used to screen multiple scaffolds against target proteins to select for high affinity binders. Scaffolds are displayed on phage particles and can be tested for binding to targets with standard binding assays. Various target proteins are encoded by essential genes to disrupt cellular proliferation and were identified as fitness genes in CRISPR screens. Altogether, utilizing the two technologies facilitates making and screening of binders to intracellular targets and accelerates the production of highly efficient synthetic binders. To develop and optimize the technique, the Sidhu lab generated 27 protein scaffold libraries that were tested against 25 target proteins and selected several variants for further analysis in mammalian cells.M.Sc
