Quantification of a Pharmacodynamic ERK End Point
in Melanoma Cell Lysates: Toward Personalized Precision Medicine
- Publication date
- Publisher
Abstract
Protein kinases are mutated or otherwise
rendered constitutively
active in numerous cancers where they are attractive therapeutic targets
with well over a dozen kinase inhibitors now being used in therapy.
While fluorescent sensors have capacity to measure changes in kinase
activity, surprisingly they have not been utilized for biomarker studies.
A first-generation peptide sensor for ERK based on the Sox fluorophore
is described. This sensor called ERK-sensor-D1 possesses high activity
toward ERK and more than 10-fold discrimination over other MAPKs.
The sensor can rapidly quantify ERK activity in cell lysates and monitor
ERK pathway engagement by BRAF and MEK inhibitors in cultured melanoma
cell lines. The dynamic range of the sensor assay allows ERK activities
that have potential for profound clinical consequences to be rapidly
distinguished