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Identification of hemoglobin variants by top-down mass spectrometry using selected diagnostic product ions

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Abstract

Hemoglobin disorder diagnosis is a complex procedure combining several analytical steps. Due to the lack of specificity of the currently used protein analysis methods, the identification of uncommon hemoglobin variants (proteoforms) can become a hard task to accomplish. The aim of this work was to develop a mass spectrometry-based approach to quickly identify mutated protein sequences within globin chain variants. To reach this goal, a top-down electron transfer dissociation mass spectrometry method was developed for hemoglobin β chain analysis. A diagnostic product ion list was established with a color code strategy allowing to quickly and specifically localize a mutation in the hemoglobin β chain sequence. The method was applied to the analysis of rare hemoglobin β chain variants and an Aγ-β fusion protein. The results showed that the developed data analysis process allows fast and reliable interpretation of top-down electron transfer dissociation mass spectra by nonexpert users in the clinical area.

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Acknowledgments

This study was supported by the Swiss National Science Foundation Grant No. 32003B_143809 and by the Ernst & Lucie Schmidheiny Foundation. The authors thank Paola Antinori, Fabienne Jeanneret, and HuiSong Pak for their scientific support and encouraging discussions.

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Authors and Affiliations

  1. Department of Human Protein Sciences, Faculty of Medicine, Geneva University, Rue Michel Servet 1, 1211, Geneva, Switzerland

    Didia Coelho Graça, Denis Hochstrasser, Alexander Scherl & Pierre Lescuyer

  2. Bruker Daltonics, Fahrenheitstraße 4, 28359, Bremen, Germany

    Ralf Hartmer, Wolfgang Jabs & Carsten Stoermer

  3. Hematology Service, Geneva University Hospitals, Rue Gabrielle-Perret-Gentil 4, 1205, Geneva, Switzerland

    Photis Beris & Kaveh Samii

  4. Laboratoire Unilabs Coppet, Chemin des Perrières 2, 1296, Coppet, Switzerland

    Photis Beris

  5. Department of Genetic and Laboratory Medicine, Geneva University Hospitals, Rue Gabrielle-Perret-Gentil 4, 1205, Geneva, Switzerland

    Lorella Clerici, Denis Hochstrasser, Alexander Scherl & Pierre Lescuyer

  6. Biomolecular Mass Spectrometry Laboratory, Ecole Polytechnique Fédérale de Lausanne, 1015, Lausanne, Switzerland

    Yury O. Tsybin

Authors
  1. Didia Coelho Graça
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  2. Ralf Hartmer
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  3. Wolfgang Jabs
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  4. Photis Beris
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  5. Lorella Clerici
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  6. Carsten Stoermer
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  7. Kaveh Samii
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  8. Denis Hochstrasser
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  9. Yury O. Tsybin
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  10. Alexander Scherl
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  11. Pierre Lescuyer
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Corresponding author

Correspondence to Pierre Lescuyer.

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Coelho Graça, D., Hartmer, R., Jabs, W. et al. Identification of hemoglobin variants by top-down mass spectrometry using selected diagnostic product ions. Anal Bioanal Chem 407, 2837–2845 (2015). https://doi.org/10.1007/s00216-015-8525-5

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  • DOI: https://doi.org/10.1007/s00216-015-8525-5

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