Targeted mass-spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP kinase, and AKT signaling

• DOI: 10.1016/j.crmeth.2021.100015
• Publication Date: 2021-06-14T14:27:03Z
• Authors: Jeffrey R. Whiteaker; Kanika Sharma; Melissa A. Hoffman; Eric Kuhn; Lei Zhao; Alexandra R. Cocco; Regine M. Schoenherr; Jacob J. Kennedy; Ulianna Voytovich; Chenwei Lin; Bin Fang; Kiah Bowers; Gordon Whiteley; Simona Colantonio; William Bocik; Rhonda Roberts; Tara Hiltke; Emily Boja; Henry Rodriguez; Frank McCormick; Matthew Holderfield; Steven A. Carr; John M. Koomen; Amanda G. Paulovich
• Keywords: Multiplex

A primary goal of the US National Cancer Institute's Ras initiative at Frederick Laboratory for Research is to develop methods quantify RAS signaling facilitate development novel cancer therapeutics. We use targeted proteomics technologies a community resource consisting 256 validated multiple reaction monitoring (MRM)-based, multiplexed assays quantifying protein expression and phosphorylation through receptor tyrosine kinase, MAPK, AKT networks. As proof concept, we response melanoma (A375 SK-MEL-2) colorectal (HCT-116 HT-29) cell lines BRAF inhibition by PLX4720. These replace over 60 western blots with quantitative mass-spectrometry-based high molecular specificity precision, showing value these pharmacodynamic measurements mechanism-of-action studies. Methods, fit-for-purpose validation, results are publicly available as assays.cancer.gov.