Rowan Matney1, Kratika Singhal1, Fang Liu1, Sarah Trusiak2, Emily Chen2, Ryan D. Leib1, Allis S. Chien1
International Hupo 2020
To tackle questions of differential protein expression in the expanding phosphoproteome, one emerging strategy is the targeted measurement of protein candidates from critical pathways. Here, we use a targeted sample preparation strategy (SureQuant) to enrich clinical tissue samples for proteins specific to the AKT/mTOR pathway. We compare results across two tissue types: frozen brain tissue and formalin-fixed paraffin-embedded (FFPE) brain tissue slides, as well as two control lysates: A549 (lung cancer) and MCF7 (breast cancer). This robust method allows for multiplexed immunoprecipitation in tandem with selective probing for pathway-relevant phosphorylated proteins. In this case, these targets are 30 unique peptides from 10 proteins in the AKT/mTOR pathway. This work is foundational for future studies incorporating both quantitation and the integration of genomic data.
1. Stanford University Mass Spectrometry, Stanford, CA
2. Thermo Fisher Precision Medicine Science Center, Cambridge, MA
United States
