Online LC-FAIMS-MS/MS for the Analysis of Phosphorylation in Proteins

  • Hongyan Zhao
  • Andrew J. Creese
  • Helen J. Cooper
Part of the Methods in Molecular Biology book series (MIMB, volume 1355)


High-field asymmetric waveform ion mobility spectrometry (FAIMS) is a gas-phase separation technique which, when coupled with liquid chromatography tandem mass spectrometry, offers benefits for analysis of complex proteomics samples such as those encountered in phosphoproteomics experiments. Results from LC-FAIMS-MS/MS are typically complementary, in terms of proteome coverage and isomer identification, to those obtained by use of solution-phase separation methods, such as prefractionation with strong cation-exchange chromatography. Here, we describe the protocol for large-scale phosphorylation analysis by LC-FAIMS-MS/MS.

Key words

Phosphorylation Ion mobility spectrometry FAIMS 



The Advion Triversa Nanomate, Dionex LC and Thermo Fisher Velos Orbitrap mass spectrometer used in this research were funded through the Birmingham Science City Translational Medicine: Experimental Medicine Network of Excellence Project, with support from Advantage West Midlands (AWM). The Chinese Scholarship Council is gratefully acknowledged for funding. HJC is an EPSRC Established Career Fellow (EP/L023490/1).


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Hongyan Zhao
    • 1
  • Andrew J. Creese
    • 1
  • Helen J. Cooper
    • 1
  1. 1.School of BiosciencesUniversity of BirminghamEdgbaston, BirminghamUK

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