Diagnosis of Hemoglobinopathy and β-Thalassemia by 21 Tesla Fourier Transform Ion Cyclotron Resonance Mass Spectrometry and Tandem Mass Spectrometry of Hemoglobin from Blood
Abstract
BACKGROUND: Hemoglobinopathies and thalassemias are the most common genetically determined disorders. Current screening methods include cation-exchange HPLC and electrophoresis, the results of which can be ambiguous because of limited resolving power. Subsequently, laborious genetic testing is required for confirmation. METHODS: We performed a top-down tandem mass spectrometry (MS/MS) approach with a fast data acquisition (3 min), ultrahigh mass accuracy, and extensive residue cleavage by use of positive electrospray ionization 21 Tesla Fourier transform ion cyclotron resonance–tandem mass spectrometry (21 T FT-ICR MS/MS) for hemoglobin (Hb) variant de novo sequencing and β-thalassemia diagnosis. RESULTS: We correctly identified all Hb variants in blind analysis of 18 samples, including the first characterization of homozygous Hb Himeji variant. In addition, an Hb heterozygous variant with isotopologue mass spacing as small as 0.0194 Da (Hb AD) was resolved in both precursor ion mass spectrum (MS1) and product ion mass spectrum (MS2). In blind analysis, we also observed that the abundance ratio between intact δ and β subunits (δ/β) or the abundance ratio between intact δ and α subunits (δ/α) could serve to diagnose β-thalassemia trait caused by a mutation in 1 HBB gene. CONCLUSIONS: We found that 21 T FT-ICR MS/MS provides a benchmark for top-down MS/MS analysis of blood Hb. The present method has the potential to be translated to lower resolving power mass spectrometers (lower field FT-ICR mass spectrometry and Orbitrap) for Hb variant analysis (by MS1 and MS2) and β-thalassemia diagnosis (MS1).
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He et al. used ultrahigh-resolution MS on a 21 T FTICR to diagnose hemoglobinopathy and β-thalassemia from blood with a 3 min assay.
“He et al. utilized ultrahigh-resolution MS on a 21 T FTICR enabling the diagnosis of hemoglobinopathy and β-thalassemia from blood with a 3 min assay [158].”
From Top-down proteomics: challenges, innovations, and applications in basic and clinical research (2020) · cites this paper as [158]
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Top-down proteomics: challenges, innovations, and applications in basic and clinical research
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- Authors
- He, L., Rockwood, A. L., Agarwal, A. M., Anderson, L. C., Weisbrod, C. R., Hendrickson, C. L., & Marshall, A. G.
- Affiliations
- Florida State University (US)
- University of Utah (US)
- Corresponding author
- Marshall, Alan G
- ORCID
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2019-08-01
- Volume
- 65
- Issue
- 8
- Pages
- 986-994
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W2942883750
- Cited by
- 43
- References
- 28
- Citations by year
- 2026: 5 · 2025: 5 · 2024: 3 · 2023: 4 · 2022: 5 · 2021: 7 · 2020: 10 · 2019: 3
- FWCI
- 3.43
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Clinical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (DMR-1,644,779, 1157490)
- Division of Materials Research (1644779, DMR- 1644779, 11‐57490, DMR 1157490)
- State of Florida
- Topics
- Health Sciences › Medicine › Genetics › Hemoglobinopathies and Related Disorders
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- hemoglobinopathies, beta-thalassemia, hemoglobin variants, beta-globin gene mutations
- MeSH terms
- Amino Acid Sequence, Fourier Analysis, Hemoglobinopathies, Hemoglobins, Humans, Sensitivity and Specificity, Mass Spectrometry, Genetic Variation, beta-Thalassemia, Cyclotrons, Sequence Analysis, Protein, Tandem Mass Spectrometry, alpha-Globins, beta-Globins, delta-Globins
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