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Single-Strand DNA Binding Proteins

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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...

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Screening for Functional Non-coding Genetic Variants Using Electrophoretic Mobility Shift Assay EMSA and DNA-affinity Precipitation Assay DAPA

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On-DNA Binder Confirmation: Increasing Confidence in DEL Hits.

Mark A Mantell¹, Karanbir S Pahil¹, Karli R Holman¹

¹Encoded Technologies, Molecular Modalities Discovery, GSK, Cambridge, Massachusetts 02140, United States.

Journal of Medicinal Chemistry

|September 10, 2025

Summary

This summary is machine-generated.

This study introduces an on-DNA workflow for DNA-encoded libraries (DELs) to improve small-molecule drug discovery. This method increases throughput and confidence in identifying drug leads by confirming binders directly on DNA, reducing failed experiments.

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Area of Science:

Medicinal Chemistry
Biotechnology
Drug Discovery

Background:

DNA-encoded libraries (DELs) are crucial for identifying small-molecule leads against protein targets.
Traditional off-DNA resynthesis and testing of DEL hits are resource-intensive and often yield false positives.

Purpose of the Study:

To develop and validate an on-DNA workflow for hit resynthesis and binder confirmation in DEL screens.
To increase throughput and reliability in identifying confirmed DEL hits for drug discovery.

Main Methods:

Implementation of GSK's on-DNA hit resynthesis and binder confirmation platform.
Utilized thermal shift, microscale thermophoresis, activity assays, and compound-immobilized SPR for initial derisking.
Employed mass spectrometry for specific binder identification in complex on-DNA mixtures.

Main Results:

The on-DNA workflow enhances throughput and emulates original library synthesis.
Successfully identified side product binders and increased confidence in DEL hits.
Demonstrated robust platform for derisking DEL hits and identifying specific binders.

Conclusions:

GSK's on-DNA binder confirmation platform significantly improves the efficiency and accuracy of DEL hit evaluation.
This approach enables effective evaluation and expansion of hits from DEL screens, accelerating drug discovery pipelines.