Selection methods for proximity-dependent enrichment of ligands from DNA-encoded libraries using enzymatic fusion proteins

Herein, we report a selection approach to enrich ligands from DNA-encoded libraries (DELs) based on proximity to an enzymatic tag on the target protein. This method involves uncaging or installation of a biotin purification tag on the DNA construct either through photodeprotection of a protected biotin group using a light emitting protein tag (nanoluciferase) or by acylation using an engineered biotin ligase (UltraID). This selection does not require purification of the target protein and results in improved recovery and enrichment of DNA-linked ligands. This approach should serve as a general and convenient tool for molecular discovery with DELs. Nanoluciferase or engineered biotin ligase fusions to a protein target allow proximity-induced biotinylation of DNA-linked ligands. The approach benefits ligand enrichment from DNA-encoded chemical libraries (DELs) and live cell selections.