|Bhavin Patel , MD. works as a senior research and development scientist in MS reagents group at Thermo Fisher Scientific. I have a medical Doctor degree from the MS University in India, and a masters in biotechnology from the Georgetown University. I had led a protein biomarker targeted MS assay group at NextGen Sciences and a proteomics/mass spectrometry group at Fox Chase Cancer Center before joining Thermo Fisher Scientific in 2013. Since joining Thermo, I have led the development of targeted MS assay kits, new MS system suitability standards, protein sample preparation reagents, antibody validation method for IP-MS, and reagents for targeted quantitative proteomic analysis. |
Targeted Mass Spectrometry Assays for Absolute Quantitation of AKT/mTOR Signaling Pathway ProteinsProteomics offers a deeper understanding of the protein events influencing cellular and biological function. However, Identifying key regulated proteins and their implicated signaling pathways represents a major analytical challenge to researchers. We have optimized a multiplex immunoprecipitation to targeted mass spectrometry (mIP-tMS) workflow to develop the SureQuant pathway panels, achieving simultaneous enrichment and absolute quantitation of multiple total and phosphorylated proteins from the AKT/mTOR pathway. Read More
We discuss improved enrichment/sample preparation method as well as development and validation of targeted MS assay including calibration curves, robustness, precision and accuracy of these assays. We have utilized the Skyline software for assay development and data analysis for absolute quantitation of AKT pathway proteins. Additionally, we have developed a system suitability standard containing isotopologue peptides mixture and utilized Skyline for data analysis. Finally, we’ll examine applications of multiplexed IP to targeted MS assays compared to existing immunoassay techniques.