title: Automated screening for small organic ligands using DNA-encoded chemical libraries.
abstract: DNA-encoded chemical libraries (DECLs) are collections of organic compounds that are individually linked to different oligonucleotides, serving as amplifiable identification barcodes. As all compounds in the library can be identified by their DNA tags, they can be mixed and used in affinity-capture experiments on target proteins of interest. In this protocol, we describe the screening process that allows the identification of the few binding molecules within the multiplicity of library members. First, the automated affinity selection process physically isolates binding library members. Second, the DNA codes of the isolated binders are PCR-amplified and subjected to high-throughput DNA sequencing. Third, the obtained sequencing data are evaluated using a C++ program and the results are displayed using MATLAB software. The resulting selection fingerprints facilitate the discrimination of binding from nonbinding library members. The described procedures allow the identification of small organic ligands to biological targets from a DECL within 10 d.
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Willy
authors.initials: W
authors.lastname: Decurtins
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Moreno
authors.initials: M
authors.lastname: Wichert
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Raphael M
authors.initials: RM
authors.lastname: Franzini
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Fabian
authors.initials: F
authors.lastname: Buller
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Stravs
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Yixin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Dario
authors.initials: D
authors.lastname: Neri
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH Zurich), Zurich, Switzerland.
authors.forename: Jörg
authors.initials: J
authors.lastname: Scheuermann
mesh_terms: D018736:DNA Adducts / Q000032:analysis*; D059014:High-Throughput Nucleotide Sequencing; D008024:Ligands; D008403:Mass Screening / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D054852:Small Molecule Libraries
chemical_list: D018736:DNA Adducts; D008024:Ligands; D009841:Oligonucleotides; D054852:Small Molecule Libraries