Targeted protein degradation (TPD) has transformed drug discovery by enabling selective elimination of disease-relevant proteins. Cereblon (CRBN) modulators, including IMiDs and next-generation molecular glues -drive targeted ubiquitination of transcription factors and other neosubstrates, shaping both therapeutic response and safety. However, traditional Western blot–based methods for profiling these degraders are low-throughput, labor-intensive, and difficult to scale.

Revvity’s no-wash HTRF™ assays provide a fast, quantitative, and highly reproducible alternative. This application note profiles six CRBN modulators—thalidomide, lenalidomide, iberdomide (CC-220), FPFT-2216, TMX-4116, and eragidomide (CC-90009)—across five key neosubstrates (IKZF1, IKZF3, CK1α, SALL4, and GSPT1) in cell-based assays.

Key highlights include: 

• Side-by-side profiling of six CRBN modulators across five key neosubstrates.
• Clear degradation signatures showing potency, maximal degradation, and time-dependent kinetics.
• Distinct selectivity patterns from IKZF-specific to CK1α or GSPT1-focused activity.
• Unexpected SALL4 degradation revealing broader neosubstrate engagement.
• Fast, no-wash HTRF workflow enabling reproducible, high-throughput endogenous protein measurement.

For research use only. Not for use in diagnostic procedures.