Biosensor-based small molecule fragment screening with biolayer interferometry.

Area of Science:

• Biochemistry
• Drug Discovery
• Biotechnology

Background:

• Biosensor-based fragment screening aids drug discovery by differentiating true hits from false positives.
• Biolayer interferometry (BLI) is an emerging technique for monitoring small molecule interactions using disposable sensors.
• BLI offers advantages over traditional biochemical methods by analyzing binding profiles and responses.

Purpose of the Study:

• To evaluate Biolayer interferometry (BLI) as a biosensor method for fragment screening.
• To assess BLI's effectiveness in identifying hits for challenging protein targets involved in protein-protein interactions (PPIs).
• To compare BLI screening results with biochemical high-throughput screening (HTS) data for the eIF4E target.

Main Methods:

• Fragment screening of 6,500 compounds using BLI on challenging targets like BCL-2, JNK1, and eIF4E.
• Validation of BLI for small molecule detection and fragment screening against model systems and well-characterized targets.
• Comparative analysis of BLI screening results with surface plasmon resonance (SPR) and biochemical HTS data.

Main Results:

• BLI demonstrated comparable affinity constants and binding profiles to SPR for model systems.
• Fragment screening using BLI identified hits for challenging PPI targets, including BCL-2, JNK1, and eIF4E.
• For eIF4E, BLI screening yielded overlapping hits with biochemical HTS for the PPI site and unique hits, validating its utility.

Conclusions:

• BLI is a valuable biosensor tool for fragment-based drug discovery, capable of reducing false-positive rates.
• BLI effectively screens challenging targets, including those involved in protein-protein interactions.
• Comparative analysis confirms BLI's utility and potential to identify novel drug leads distinct from traditional methods.