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Monitoring of the AKT/mTOR pathway using Internal Standard Triggered-Parallel Reaction Monitoring (IS-PRM)

Reputable Mentor II
Reputable Mentor II
Shou_Ling Xu1; Michael Blank1; Sebastien Gallien2; Bhavin Patel3; Alex Behling3; Leigh Foster3; John Rogers3; Bruno Domon2; Andreas Huhmer1

Monitoring multiple proteins with high precision is essential to biological and biomedical sciences. Targeted proteomics techniques, such as Parallel Reaction Monitoring (PRM), have enabled the confident quantification of sets of proteins with high sensitivity and selectivity. In this study, an advanced data acquisition method, called IS-PRM, was applied to quantify large sets of peptides in complex matrices. This method relies on monitoring the internal standards added to the samples, while adjusting on-the-fly the elution windows of the targeted peptides to improve the acquisition efficiency. The method was applied to the analysis of proteins involved in AKT/mTOR pathway with synthetic peptide standards mixtures in a matrix of six proteins and demonstrated the sensitive and confident detection and quantification of proteins. We will implement this method with complex matrices in five cancel cell lysates in the next step.

1Thermo Fisher Scientific, San Jose, CA 2Luxembourg Clinical Proteomics Center, Strassen, Luxembourg 3Thermo Fisher Scientific, Rockford, IL
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Last update:
‎10-15-2021 11:33 AM
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