Multiplexed liquid chromatography-multiple reaction monitoring mass spectrometry quantification of cancer signaling proteins

Yi Chen, Kate J. Fisher, Mark Lloyd, Elizabeth R. Wood, Domenico Coppola, Erin Siegel, David Shibata, Yian A. Chen, John M. Koomen

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages19-45
Number of pages27
DOIs
StatePublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1647
ISSN (Print)1064-3745

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Liquid Chromatography
Mass Spectrometry
Neoplasms
Proteins
Cell Line
Catenins
Wnt Signaling Pathway
Mitogen-Activated Protein Kinase Kinases
Colonic Neoplasms
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Digestion
Apoptosis
Peptides
Therapeutics

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Chen, Y., Fisher, K. J., Lloyd, M., Wood, E. R., Coppola, D., Siegel, E., ... Koomen, J. M. (2017). Multiplexed liquid chromatography-multiple reaction monitoring mass spectrometry quantification of cancer signaling proteins. In Methods in Molecular Biology (pp. 19-45). (Methods in Molecular Biology; Vol. 1647). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7201-2_2

Multiplexed liquid chromatography-multiple reaction monitoring mass spectrometry quantification of cancer signaling proteins. / Chen, Yi; Fisher, Kate J.; Lloyd, Mark; Wood, Elizabeth R.; Coppola, Domenico; Siegel, Erin; Shibata, David; Chen, Yian A.; Koomen, John M.

Methods in Molecular Biology. Humana Press Inc., 2017. p. 19-45 (Methods in Molecular Biology; Vol. 1647).

Research output: Chapter in Book/Report/Conference proceedingChapter

Chen, Y, Fisher, KJ, Lloyd, M, Wood, ER, Coppola, D, Siegel, E, Shibata, D, Chen, YA & Koomen, JM 2017, Multiplexed liquid chromatography-multiple reaction monitoring mass spectrometry quantification of cancer signaling proteins. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1647, Humana Press Inc., pp. 19-45. https://doi.org/10.1007/978-1-4939-7201-2_2
Chen Y, Fisher KJ, Lloyd M, Wood ER, Coppola D, Siegel E et al. Multiplexed liquid chromatography-multiple reaction monitoring mass spectrometry quantification of cancer signaling proteins. In Methods in Molecular Biology. Humana Press Inc. 2017. p. 19-45. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7201-2_2
Chen, Yi ; Fisher, Kate J. ; Lloyd, Mark ; Wood, Elizabeth R. ; Coppola, Domenico ; Siegel, Erin ; Shibata, David ; Chen, Yian A. ; Koomen, John M. / Multiplexed liquid chromatography-multiple reaction monitoring mass spectrometry quantification of cancer signaling proteins. Methods in Molecular Biology. Humana Press Inc., 2017. pp. 19-45 (Methods in Molecular Biology).
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