Quantitative Mass Spectrometry Analysis of Intact Hemoglobin A<sub>2</sub> by Precursor Ion Isolation and Detection

Acosta‐Martin, Adelina E.; Graca, Didia Coelho; Antinori, Paola; Clerici, Lorella; Hartmer, Ralf; Meyer, Markus R.; Hochstrasser, Denis F.; Samii, Kaveh; Lescuyer, Pierre; Scherl, Alexander

doi:10.1021/ac401782t

Cited by 14 publications

(10 citation statements)

References 8 publications

(9 reference statements)

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“…A step in this direction was taken recently by developing a top-down assay to monitor clinically relevant hemoglobin variants caused by single nucleotide polymorphisms at the level of the fragment ion (72). Previously, a clinically applicable ion trap assay was developed to quantify hemoglobin A2 variants at the MS 1 level to aid thalassemia diagnosis, and it demonstrated impressive analytical precision that would be acceptable in diagnostic laboratories (73). Opportunities for proteoform-resolved measurements in translational research and clinical practice will continue to become available as the quantitative potential of targeted top-down measurement improves—most likely in conjunction with advances in instrument speed, precursor activation, and fragment ion detection.…”

Section: Paths Forward In Top-down Proteomicsmentioning

confidence: 99%

Progress in Top-Down Proteomics and the Analysis of Proteoforms

Toby

Fornelli

Kelleher

2016

Annual Rev. Anal. Chem.

467

456

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From a molecular perspective, enactors of function in biology are intact proteins that can be variably modified at the genetic, transcriptional, or post-translational level. Over the past 30 years, mass spectrometry (MS) has become a powerful method for the analysis of proteomes. Prevailing bottom-up proteomics operates at the level of the peptide, leading to issues with protein inference, connectivity, and incomplete sequence/modification information. Top-down proteomics (TDP), alternatively, applies MS at the proteoform level to analyze intact proteins with diverse sources of intramolecular complexity preserved during analysis. Fortunately, advances in prefractionation workflows, MS instrumentation, and dissociation methods for whole-protein ions have helped TDP emerge as an accessible and potentially disruptive modality with increasingly translational value. In this review, we discuss technical and conceptual advances in TDP, along with the growing power of proteoform-resolved measurements in clinical and translational research.

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Section: Paths Forward In Top-down Proteomicsmentioning

confidence: 99%

Progress in Top-Down Proteomics and the Analysis of Proteoforms

Toby

Fornelli

Kelleher

2016

Annual Rev. Anal. Chem.

467

456

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“…3 ). Moreover, the Hb-E variant is known to cause weak union between α- and β-globin that leads to fast elution of these globin chains within the first minute as a free globin that causes higher amplitudes of the unknown HPLC peaks particularly in patients with Hb-E B-Thal type [ 8 , [17] , [18] , [19] ].…”

Section: Discussionmentioning

confidence: 99%

Relative proteome quantification of alpha, beta, gamma and delta globin chains in early eluting peaks of Bio-Rad variant II® CE-HPLC of hemoglobin from healthy and beta-thalassemia subjects in Malaysia

Abdullah

Ibrahim

Jassim

et al. 2019

Biochemistry and Biophysics Reports

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This is the first report of QQQ-mass spectrometric identification and quantification of the Hb subunits, alpha, beta, delta and gamma globin peptides, derived from enzymatic-digestion of proteins in the early unknown peaks of the Bio-Rad cation-exchange chromatography of haemoglobin. The objectives were to assess the relationship of the quantity of the free alpha, beta, delta and gamma globin chains with the phenotypic diversity of beta-thalassaemias (β-thal). The results demonstrate that the pools of free globin chains in red blood cells were correlating with the severity of the disease in patients with different phenotypes of β-thal. The mechanism and the regulation of synthesis of free globin chains pool in a normal individual and in patients with different β-thal phenotypes could arise from several mechanisms which will require further investigation. The role of the free globin pool in patients with β-thal for development of novel therapeutic approaches based on these potential targets requires further investigation. Pertinent biomarkers improves the diagnosis of the β-thal, especially in low-income countries where they are most common and allows more effective therapeutic intervention leading to more successful therapeutic outcome.

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“…Results from DBS spots were similar to those obtained using the standard sample preparation procedure described in the article (data not shown). Finally, this top-down ETD MS method is complementing our previously described MS methods for the identification of common Hb variants and quantification of Hb chains [13,22].…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2015

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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 (A)γ-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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Quantitative Mass Spectrometry Analysis of Intact Hemoglobin A₂ by Precursor Ion Isolation and Detection

Cited by 14 publications

References 8 publications

Progress in Top-Down Proteomics and the Analysis of Proteoforms

Progress in Top-Down Proteomics and the Analysis of Proteoforms

Relative proteome quantification of alpha, beta, gamma and delta globin chains in early eluting peaks of Bio-Rad variant II® CE-HPLC of hemoglobin from healthy and beta-thalassemia subjects in Malaysia

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

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Quantitative Mass Spectrometry Analysis of Intact Hemoglobin A2 by Precursor Ion Isolation and Detection

Cited by 14 publications

References 8 publications

Progress in Top-Down Proteomics and the Analysis of Proteoforms

Progress in Top-Down Proteomics and the Analysis of Proteoforms

Relative proteome quantification of alpha, beta, gamma and delta globin chains in early eluting peaks of Bio-Rad variant II® CE-HPLC of hemoglobin from healthy and beta-thalassemia subjects in Malaysia

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

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Quantitative Mass Spectrometry Analysis of Intact Hemoglobin A₂ by Precursor Ion Isolation and Detection