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A Protocol for Top-Down Proteomics Using HPLC and ETD/PTR-MS

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LC-MS/MS in Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 658))

Abstract

Analysis of intact proteins by tandem mass spectrometry has mostly been confined to high-end mass spectrometry platforms. This protocol describes the application of routine HPLC to separate proteins, MALDI-ToF mass spectrometry to interrogate intact protein species and electron transfer dissociation/proton transfer reaction within a quadrupole ion trap to perform tandem mass spectrometry.

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Acknowledgements

I would like to thank current and former members of the Michael Barber Centre, University of Manchester for constructive comments during this work. John Cottrell (Matrix Sciences) provided demonstration licensing of top-down Mascot. Ken Cook (Dionex) provided demonstration of HPLC columns and assisted in setting up gradients. Carsten Baessmann, Andrea Kiehne, Markus Lubeck, Andrea Schneider and Julia Smith (Bruker Daltonics) have provided essential guidance both with ETD/PTR experiments and with data processing.

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Authors and Affiliations

  1. Michael Barber Centre for Mass Spectrometry, University of Manchester, Manchester, UK

    Sarah R. Hart

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  1. Sarah R. Hart
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Editor information

Editors and Affiliations

  1. Bart’s and the London School of Medicine, Institute of Cancer, Queen Mary, Universtiy of London, Charterhouse Square, London, EC1M 6BQ, United Kingdom

    Pedro R. Cutillas

  2. , EGA Institute for Women's Health, University College London, Gower Street, London, WC1E 6BT, United Kingdom

    John F. Timms

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Hart, S.R. (2010). A Protocol for Top-Down Proteomics Using HPLC and ETD/PTR-MS. In: Cutillas, P., Timms, J. (eds) LC-MS/MS in Proteomics. Methods in Molecular Biology, vol 658. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-780-8_21

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  • DOI: https://doi.org/10.1007/978-1-60761-780-8_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-779-2

  • Online ISBN: 978-1-60761-780-8

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