Promega, N8012, NanoBRET® TE Intracellular RAS Assay

Measure Orthosteric and Allosteric RAS Ligands Across Multiple RAS Variants
The NanoBRET® TE Intracellular RAS Assay can be used to measure compound cellular affinity and occupancy for multiple RAS proteins, including wildtype and mutant KRAS/HRAS. The assay can detect compound binding to both switch I/II pocket and switch II pocket.
Intracellular engagement of orthosteric and allosteric inhibitors for KRAS(G12C). The NanoBRET® TE RAS Assay was used to quantify the cellular affinity of RAS inhibitors in HEK293 cells expressing SmBiT(q)-KRAS(G12C) and LgBiT-KRAS(G12C) Fusion Vectors. Apparent affinity was measured for the switch I/II pocket binder BI-2852, for covalent switch II pocket binder Sotorasib and Adagrasib, and for non-covalent switch II pocket binder MRTX1133.
Compound selectivity across RAS variants. The NanoBRET® TE RAS Assay allows selectivity evaluation of test compounds across multiple RAS variants using the same workflow and a consistent tracer concentration of 2µM. Shown here are examples of selectivity profiles for Sotorasib ( Panel A ) and MRTX1133 ( Panel B ) across the entire suite of available RAS fusion vectors. Sotorasib demonstrates strong selectivity for G12C hotspot RAS mutants, while MRTX1133 potently engages its intended target KRAS(G12D) and many other KRAS variants.