DNA-encoded synthesis can generate vastly diverse screening libraries of arbitrarily complex molecules as long chemical reaction conditions do not compromise DNA's informational integrity, a fundamental constraint that "DNA-compatible" development does presently address. We devised rehearsal, an integrated analysis yield and impact on DNA, to acquire these key missing data. Magnetic DNA-functionalized sensor beads quantitatively report the % DNA template remaining viable for PCR...

High throughput DNA sequencing has been performed by using a microfabricated 96-channel radial capillary array electrophoresis (μCAE) microchannel plate detected 4-color rotary confocal fluorescence scanner. The features novel injector for uniform sieving matrix loading as well high resolution, tapered turns that provide an effective separation length of 15.9 cm on compact 150-mm diameter wafer. Expanded common buffer chambers the cathode, anode, and waste reservoirs are used to simplify...

Among the molecular milieu of cell, membrane bilayer stands out as a complex and elusive synthetic target. We report microfluidic assembly line that produces uniform cellular compartments from droplet, lipid, oil/water interface starting materials. Droplets form in lipid-containing oil flow travel to junction where confluence extracellular aqueous media establishes flow-patterned is both stable reproducible. A triangular post mediates phase transfer by deflecting droplets oil, through...

The promise of exploiting combinatorial synthesis for small molecule discovery remains unfulfilled due primarily to the "structure elucidation problem": back-end mass spectrometric analysis that significantly restricts one-bead-one-compound (OBOC) library complexity. very molecular features confer binding potency and specificity, such as stereochemistry, regiochemistry, scaffold rigidity, are conspicuously absent from most libraries because isomerism introduces redundancy diverse scaffolds...

Synthetic biology aims to improve human health and the environment by repurposing biological enzymes for use in practical applications. However, natural often function with suboptimal activity when engineered into pathways or challenged recognize unnatural substrates. Overcoming this problem requires efficient directed evolution methods discovering new enzyme variants that a desired activity. Here, we describe construction, validation, application of fluorescence-activated droplet sorting...

Significance Drug discovery generally investigates one target at a time, in sharp contrast to living organisms, which mold ligands and targets by evolution of highly complex molecular interaction networks. We recapitulate this modality encoding drug structures DNA, allowing the entire DNA-encoded library interact with thousands RNA fold targets, then decoding both sequencing. This information serves as filter identify human RNAs aberrantly produced cancer that are also binding partners...

Turns in microfabricated capillary electrophoresis channels generally result degraded separation quality. To circumvent this limitation, were constructed with different types of turns to determine the design that minimizes turn-induced band broadening. In particular, tapered created by narrowing channel width before start a turn and widening after is complete. The radius curvature turn, length over which tapered, degree tapering explored. column efficiencies determined examining resolution...

Hydrogenated amorphous silicon (a-Si:H) PIN photodiodes have been developed and characterized as fluorescence detectors for microfluidic analysis devices. A discrete a-Si:H photodiode is first fabricated on a glass substrate used to detect fluorescent dye standards using conventional confocal microscopy. In this format, the limit of detection fluorescein flowing in 50-microm deep channel 680 pM (S/N = 3). hybrid integrated system consisting half-ball lens, ZnS/YF3 multilayer optical...

DNA-encoded synthesis is rekindling interest in combinatorial compound libraries for drug discovery and technology automated quantitative library screening. Here, we disclose a microfluidic circuit that enables functional screens of beads. The device carries out bead distribution into picoliter-scale assay reagent droplets, photochemical cleavage from the bead, incubation, laser-induced fluorescence-based detection, fluorescence-activated droplet sorting to isolate hits. beads (10-μm...

Robotic high-throughput compound screening (HTS) and, increasingly, DNA-encoded library (DEL) are driving bioactive chemical matter discovery in the postgenomic era. HTS enables activity-based investigation of highly complex targets using static libraries. Conversely, DEL grants efficient access to novel diversity, although is limited affinity-based selections. Here, we describe an integrated droplet-based microfluidic circuit that directly screens solid-phase DELs for activity. An example...

ConspectusThe Human Genome Project ultimately aimed to translate DNA sequence into drugs. With the draft in hand, Molecular Libraries Program set out prosecute all genome-encoded proteins for drug discovery with automated high-throughput screening (HTS). This ambitious vision remains unfulfilled, even while innovations sequencing technology have fully democratized access genome-scale sequencing. Why? While central dogma of biology allows us chart entirety cellular metabolism through...

A microfabricated electrophoretic bioprocessor for integrated DNA sequencing sample desalting, template removal, preconcentration, and CE analysis is presented. low-viscosity gel capture matrix, containing an acrylamide−copolymerized oligonucleotide complementary to the 20-base sequence directly 3' of M13−40 universal forward priming site, introduced into 60-nL chamber. Unpurified reaction products are electrophoretically driven through chamber; extension hybridize while contaminating...

The circulating antibody repertoire encodes a patient's health status and pathogen exposure history, but identifying antibodies with diagnostic potential usually requires knowledge of the antigen(s). We previously circumvented this problem by screening libraries bead-displayed small molecules against case control serum samples to discover "epitope surrogates" (ligands IgGs enriched in sample). Here, we describe an improved version technology that employs DNA-encoded high-throughput...

Preparative reactions that occur efficiently under dilute, buffered, aqueous conditions in the presence of biomolecules find application ligation, peptide synthesis, and polynucleotide synthesis sequencing. However, identification functional groups or reagents are mutually reactive with one another, but unreactive biopolymers water, is challenging. Shown here cobalt catalysts react alkenes aqueous, buffered promote efficient cycloisomerization formal Friedel-Crafts reactions. The...

DNA-encoded chemical library (DECL) synthesis must occur in aqueous media under conditions that preserve the integrity of DNA encoding tag. While identification "DNA-compatible" reaction is critical for development DECL designs explore previously inaccessible space, reports measuring such compatibility have been largely restricted to methods do not faithfully capture impact on fidelity solution phase. Here we report a comprehensive methodology uses soluble substrates exactly recapitulate...

A solid-phase DNA-encoded library (DEL) was studied for binding the RNA repeat expansion r(CUG)exp, causative agent of most common form adult-onset muscular dystrophy, myotonic dystrophy type 1 (DM1). variety uncharged and novel binders were identified to selectively bind r(CUG)exp by using a two-color flow cytometry screen. The cellular activity one binder augmented attaching it with module that directly cleaves r(CUG)exp. In DM1 patient-derived muscle cells, compound specifically bound...

DNA-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 DNA-tagged members that are ligands purified and immobilized protein target. In these selections, DNA tags can materially undesirably influence target binding and, therefore, experiment outcome. Here, we use solid-phase droplet-based microfluidic screening to separate...