co9b00153_si_001.pdf (1.95 MB)
Off-DNA DNA-Encoded Library Affinity Screening
journal contribution
posted on 2019-12-31, 12:40 authored by Amber
L. Hackler, Forrest G. FitzGerald, Vuong Q. Dang, Alexander L. Satz, Brian M. PaegelDNA-encoded library (DEL) technology is emerging as a
key element of the small molecule discovery toolbox. Conventional
DEL screens (i.e., on-DNA screening) interrogate large combinatorial
libraries via affinity selection of DNA-tagged library members that
are ligands of a purified and immobilized protein target. In these
selections, the DNA tags can materially and undesirably influence
target binding and, therefore, the experiment outcome. Here, we use
a solid-phase DEL and droplet-based microfluidic screening to separate
the DEL member from its DNA tag (i.e., off-DNA screening), for subsequent
in-droplet laser-induced fluorescence polarization (FP) detection
of target binding, obviating DNA tag interference. Using the receptor
tyrosine kinase (RTK) discoidin domain receptor 1 (DDR1) as a proof-of-concept
target in a droplet-scale competition-binding assay, we screened a
67 100-member solid-phase DEL of drug-like small molecules
for competitive ligands of DDR1 and identified several known RTK inhibitor
pharmacophores, including azaindole- and quinazolinone-containing
monomers. Off-DNA DEL affinity screening with FP detection is potentially
amenable to a wide array of target classes, including nucleic acid
binding proteins, proteins that are difficult to overexpress and purify,
or targets with no known activity assay.
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target classesOff-DNA DNA-Encoded Library Affinity Screening DNA-encoded libraryin-droplet laser-induced fluorescence polarizationdroplet-based microfluidic screeningacid binding proteinsDDR 1target bindingreceptor tyrosine kinasemolecule discovery toolboxactivity assaycombinatorial librariesDNA tagsRTK inhibitor pharmacophores67 100-discoidin domain receptor 1droplet-scale competition-binding assayinfluence target bindingFP detectionDEL memberDNA tagprotein targetproof-of-concept targetobviating DNA tag interferenceOff-DNA DEL affinity screeningquinazolinone-containing monomers67 100- member solid-phase DELsolid-phase DELConventional DEL screensaffinity selectionDNA-tagged library membersexperiment outcome
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