Category: Micro- and Nanotechnologies
DNA-encoded libraries provide an efficient and vast source of diversity for screening and are often deployed against targets that are unsuited for, or have failed to produce ligands using traditional HTS style screens. An HIV-1 protease inhibitor library, composed of variations on known FDA-approved inhibitor themes, was designed to screen for inhibitors of 6 drug resistant HIV-1 protease mutants in order to generate pan-library structure activity relationship profiles against a rapidly mutating viral target. The 29k member library was synthesized using DNA-encoded solid-phase synthesis and screened using microfluidic droplet-based functional assays. Library beads were first encapsulated into assay reagent containing droplets, compound was then photochemically dosed from the beads via irradiation of UV-cleavable photolabile linker, droplets were incubated, and finally sorted based on fluorescence. Hit beads were collected, the DNA encoding tags amplified by PCR, and the resulting amplicons were sequenced in bulk. The next-generation sequencing output was used to elucidate hit structures and prioritize hits for resynthesis based on homology and redundancy. DNA-encoded solid-phase combinatorial libraries and miniaturizing high-throughput screening provides a distributable and economical platform for small molecule discovery and facilitates efficient exploration of unconventional targets and chemical spaces.
Marie Malone– Graduate Student, The Scripps Research Institute, Jupiter, FL
The Scripps Research Institute
Marie Malone is a graduate PhD student at the Scripps Research Institute, Jupiter, FL. She received her bachelors of science from the University of Victoria, BC, Canada. She is doing her graduate work on solid-phase DNA-encoded synthesis development and it's incorporation into microfluidic droplet-based screening under Professor Brian Paegel.