On-DNA Binder Confirmation: Increasing Confidence in DEL Hits

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.