Chapter title |
Multiplexed Liquid Chromatography-Multiple Reaction Monitoring Mass Spectrometry Quantification of Cancer Signaling Proteins
|
---|---|
Chapter number | 2 |
Book title |
Proteomics for Drug Discovery
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7201-2_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7200-5, 978-1-4939-7201-2
|
Authors |
Yi Chen, Kate J. Fisher, Mark Lloyd, Elizabeth R. Wood, Domenico Coppola, Erin Siegel, David Shibata, Yian A. Chen, John M. Koomen, Chen, Yi, Fisher, Kate J., Lloyd, Mark, Wood, Elizabeth R., Coppola, Domenico, Siegel, Erin, Shibata, David, Chen, Yian A., Koomen, John M. |
Abstract |
Quantitative evaluation of protein expression across multiple cancer-related signaling pathways (e.g., Wnt/β-catenin, TGF-β, receptor tyrosine kinases (RTK), MAP kinases, NF-κB, and apoptosis) in tumor tissues may enable the development of a molecular profile for each individual tumor that can aid in the selection of appropriate targeted cancer therapies. Here, we describe the development of a broadly applicable protocol to develop and implement quantitative mass spectrometry assays using cell line models and frozen tissue specimens from colon cancer patients. Cell lines are used to develop peptide-based assays for protein quantification, which are incorporated into a method based on SDS-PAGE protein fractionation, in-gel digestion, and liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM/MS). This analytical platform is then applied to frozen tumor tissues. This protocol can be broadly applied to the study of human disease using multiplexed LC-MRM assays. |
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