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Multiplexed Small‐Molecule‐Ligand Binding Assays by Affinity Labeling and DNA Sequence Analysis

Cai, Bo ; Krusemark, Casey J.

Angewandte Chemie (International ed.), 2022-01, Vol.61 (3), p.e202113515-n/a [Periódico revisado por pares]

Germany: Wiley Subscription Services, Inc

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Título:
Multiplexed Small‐Molecule‐Ligand Binding Assays by Affinity Labeling and DNA Sequence Analysis
Autor: Cai, Bo ; Krusemark, Casey J.
Assuntos: Affinity ; Affinity labelling ; Affinity Labels - chemistry ; Assaying ; Binding ; Deoxyribonucleic acid ; DNA ; DNA - chemistry ; DNA - metabolism ; drug discovery ; Fluorescent Dyes - chemistry ; Gene sequencing ; High-Throughput Screening Assays - methods ; Labeling ; Labelling ; ligand binding assays ; Ligands ; Nucleotide sequence ; Protein Binding ; Proteins ; Proteins - chemistry ; Proteins - metabolism ; protein–ligand interactions ; Sequence analysis ; Sequence Analysis, DNA - methods ; Small Molecule Libraries - chemistry ; Small Molecule Libraries - metabolism
É parte de: Angewandte Chemie (International ed.), 2022-01, Vol.61 (3), p.e202113515-n/a
Notas: In memory of Dr. Peter J. Belshaw
The previous version of the manuscript was posted on ChemRxiv at https://chemrxiv.org/engage/chemrxiv/article‐details/60c75739f96a00132b288c20
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A previous version of this manuscript has been deposited on a preprint server
Descrição: Small‐molecule binding assays to target proteins are a core component of drug discovery and development. While a number of assay formats are available, significant drawbacks still remain in cost, sensitivity, and throughput. To improve assays by capitalizing on the power of DNA sequence analysis, we have developed an assay method that combines DNA encoding with split‐and‐pool sample handling. The approach involves affinity labeling of DNA‐linked ligands to a protein target. Critically, the labeling event assesses ligand binding and enables subsequent pooling of several samples. Application of a purifying selection on the pool for protein‐labeled DNAs allows detection of ligand binding by quantification of DNA barcodes. We demonstrate the approach in both ligand displacement and direct binding formats and demonstrate its utility in determination of relative ligand affinity, profiling ligand specificity, and high‐throughput small‐molecule screening. The affinity labeling of DNA‐barcoded ligands to target proteins allows ligand‐binding assays using DNA as a label. Pooled sample manipulation and DNA sequence detection enabled by DNA encoding yields quantitative binding assays amenable to both ligand displacement and direct binding formats.
Editor: Germany: Wiley Subscription Services, Inc
Idioma: Inglês

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Multiplexed Small‐Molecule‐Ligand Binding Assays by Affinity Labeling and DNA Sequence Analysis

Cai, Bo ; Krusemark, Casey J.

Germany: Wiley Subscription Services, Inc

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