Identification of hemoglobin variants by top-down mass spectrometry using selected diagnostic product ions
Coelho Graça, Didia ; Hartmer, Ralf ; Jabs, Wolfgang ; Beris, Photis ; Clerici, Lorella ; Stoermer, Carsten ; Samii, Kaveh ; Hochstrasser, Denis ; Tsybin, Yury ; Scherl, Alexander ; Lescuyer, Pierre
In: Analytical and Bioanalytical Chemistry, 2015, vol. 407, no. 10, p. 2837-2845
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- 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.