Rapid phenotype hemoglobin screening by high-resolution mass spectrometry on intact proteins.

Autor: Helmich F; Laboratory of Chemical Biology, Department of Biomedical Engineering, Institute of Complex Molecular Systems, Eindhoven University of Technology, Expert Center Clinical Chemistry Eindhoven, The Netherlands; Laboratory for Clinical Chemistry and Hematology, Máxima Medisch Centrum, Veldhoven, Expert Center Clinical Chemistry Eindhoven, The Netherlands. Electronic address: florishelmich@gmail.com., van Dongen JL; Laboratory of Chemical Biology, Department of Biomedical Engineering, Institute of Complex Molecular Systems, Eindhoven University of Technology, Expert Center Clinical Chemistry Eindhoven, The Netherlands., Kuijper PH; Laboratory for Clinical Chemistry and Hematology, Máxima Medisch Centrum, Veldhoven, Expert Center Clinical Chemistry Eindhoven, The Netherlands., Scharnhorst V; Expert Center Clinical Chemistry Eindhoven, Clinical Laboratory, Catharina Hospital, Eindhoven, The Netherlands., Brunsveld L; Laboratory of Chemical Biology, Department of Biomedical Engineering, Institute of Complex Molecular Systems, Eindhoven University of Technology, Expert Center Clinical Chemistry Eindhoven, The Netherlands., Broeren MA; Laboratory for Clinical Chemistry and Hematology, Máxima Medisch Centrum, Veldhoven, Expert Center Clinical Chemistry Eindhoven, The Netherlands.
Jazyk: angličtina
Zdroj: Clinica chimica acta; international journal of clinical chemistry [Clin Chim Acta] 2016 Sep 01; Vol. 460, pp. 220-6. Date of Electronic Publication: 2016 Jul 11.
DOI: 10.1016/j.cca.2016.07.006
Abstrakt: Background: Given the excellent performance of modern mass spectrometers, their clinical application for the analysis of macromolecules is a growing field of interest. This principle is explored by hemoglobin analysis, which is a representative example by its molecular weight and clinical relevance in e.g. screening programs for thalassemia and hemoglobin variants. Considering its abundance and cellular containment, pre-analysis is significantly reduced allowing for essential rapid acquisitions.
Methods: By parallel analysis of routine diagnostics for hemoglobin variants and thalassemia, we acquired samples of adults who were consented for hemoglobinopathy screening in our clinical laboratory. The pre-analytical process comprised of red cell lysis only; without further digestion and purification steps, the samples were directly injected in an electrospray ionization quadrupole time-of-flight setup and the intact proteins were analyzed by flow injection analysis. After optimization of process parameters, the deconvoluted mass spectra revealed the presence of α- and β-globulins. The reference ranges for the average mass of both globulins and their intensity ratio (α/β-ratio) were deduced from a disease-free subgroup and patients with a hemoglobinopathy were compared.
Results: The α/β-ratio is a poor marker for thalassemia patients, yet deviant α/β-ratios are found for patients with a hemoglobin variant. Mass deviations down to 1Da can be resolved; even if the patient suffers from a heterozygotic disorder, the average mass is found outside the established reference interval.
Conclusions: Although subjects with mild thalassemia were not detected, all patients with a hemoglobin variant were resolved by top-down mass spectrometry using the average globulin mass and the α/β-ratio as screening parameters.
(Copyright © 2016 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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