Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?

From its discovery, hepcidin has generated many hopes in terms of diagnosis and management of a wide variety of iron-related diseases. However, in clinical use its accurate quantification remains a challenge due to the limited sensitivity, specificity or reproducibility of the techniques described....

Full description

Bibliographic Details
Main Authors: Constance Delaby, Jérôme Vialaret, Pauline Bros, Audrey Gabelle, Thibaud Lefebvre, Hervé Puy, Christophe Hirtz, Sylvain Lehmann
Format: Article
Language:English
Published: Elsevier 2014-06-01
Series:EuPA Open Proteomics
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2212968514000087
id doaj-f02e2c48a2384a778557fb5812e52ce8
record_format Article
spelling doaj-f02e2c48a2384a778557fb5812e52ce82020-11-24T20:59:20ZengElsevierEuPA Open Proteomics2212-96852014-06-013C606710.1016/j.euprot.2014.02.004Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?Constance Delaby0Jérôme Vialaret1Pauline Bros2Audrey Gabelle3Thibaud Lefebvre4Hervé Puy5Christophe Hirtz6Sylvain Lehmann7CHU Montpellier, Institut de Recherche en Biothérapie, Hôpital St Eloi, Laboratoire de Biochimie Protéomique Clinique et CCBHM, Montpellier F-34000, FranceCHU Montpellier, Institut de Recherche en Biothérapie, Hôpital St Eloi, Laboratoire de Biochimie Protéomique Clinique et CCBHM, Montpellier F-34000, FranceCHU Montpellier, Institut de Recherche en Biothérapie, Hôpital St Eloi, Laboratoire de Biochimie Protéomique Clinique et CCBHM, Montpellier F-34000, FranceCHU Montpellier, Institut de Recherche en Biothérapie, Hôpital St Eloi, Laboratoire de Biochimie Protéomique Clinique et CCBHM, Montpellier F-34000, FranceCentre de Recherche sur l’Inflammation (CRI)/UMR 1149 INSERM – Université Paris Diderot, FranceCentre de Recherche sur l’Inflammation (CRI)/UMR 1149 INSERM – Université Paris Diderot, FranceCHU Montpellier, Institut de Recherche en Biothérapie, Hôpital St Eloi, Laboratoire de Biochimie Protéomique Clinique et CCBHM, Montpellier F-34000, FranceCHU Montpellier, Institut de Recherche en Biothérapie, Hôpital St Eloi, Laboratoire de Biochimie Protéomique Clinique et CCBHM, Montpellier F-34000, FranceFrom its discovery, hepcidin has generated many hopes in terms of diagnosis and management of a wide variety of iron-related diseases. However, in clinical use its accurate quantification remains a challenge due to the limited sensitivity, specificity or reproducibility of the techniques described. In this work, we adapted a highly specific and quantitative mass spectrometry method based on selected reaction monitoring (SRM) to measure hepcidin. Our objective was to adapt the feasibility and reproducibility of the workflow to a clinical environment. Analytical validation was performed according to ISO 15189 norms for determining the limit of detection (LOD, 2 ng/mL), limit of quantification (LOQ, 6 ng/mL), repeatability, reproducibility and linearity (up to 200 ng/mL). Using the serum of patients with various iron-related diseases we compared our SRM detection method to the well-characterized competitive ELISA (cELISA) test. The two methods were commutable (Bland–Altman plot) and we found a positive and significant correlation (r2 = 0.96, Pearson correlation coefficient p < 0.001) between both methods, although the absolute concentration determined is different from factor 5. The validation of our SRM method encourages us to propose it as an alternative approach for accurate determination of hepcidin in human samples for clinical diagnosis, follow-up and management of iron-related diseases.http://www.sciencedirect.com/science/article/pii/S2212968514000087HepcidinIron deficiencyMass spectrometryELISA
collection DOAJ
language English
format Article
sources DOAJ
author Constance Delaby
Jérôme Vialaret
Pauline Bros
Audrey Gabelle
Thibaud Lefebvre
Hervé Puy
Christophe Hirtz
Sylvain Lehmann
spellingShingle Constance Delaby
Jérôme Vialaret
Pauline Bros
Audrey Gabelle
Thibaud Lefebvre
Hervé Puy
Christophe Hirtz
Sylvain Lehmann
Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?
EuPA Open Proteomics
Hepcidin
Iron deficiency
Mass spectrometry
ELISA
author_facet Constance Delaby
Jérôme Vialaret
Pauline Bros
Audrey Gabelle
Thibaud Lefebvre
Hervé Puy
Christophe Hirtz
Sylvain Lehmann
author_sort Constance Delaby
title Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?
title_short Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?
title_full Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?
title_fullStr Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?
title_full_unstemmed Clinical measurement of Hepcidin-25 in human serum: Is quantitative mass spectrometry up to the job?
title_sort clinical measurement of hepcidin-25 in human serum: is quantitative mass spectrometry up to the job?
publisher Elsevier
series EuPA Open Proteomics
issn 2212-9685
publishDate 2014-06-01
description From its discovery, hepcidin has generated many hopes in terms of diagnosis and management of a wide variety of iron-related diseases. However, in clinical use its accurate quantification remains a challenge due to the limited sensitivity, specificity or reproducibility of the techniques described. In this work, we adapted a highly specific and quantitative mass spectrometry method based on selected reaction monitoring (SRM) to measure hepcidin. Our objective was to adapt the feasibility and reproducibility of the workflow to a clinical environment. Analytical validation was performed according to ISO 15189 norms for determining the limit of detection (LOD, 2 ng/mL), limit of quantification (LOQ, 6 ng/mL), repeatability, reproducibility and linearity (up to 200 ng/mL). Using the serum of patients with various iron-related diseases we compared our SRM detection method to the well-characterized competitive ELISA (cELISA) test. The two methods were commutable (Bland–Altman plot) and we found a positive and significant correlation (r2 = 0.96, Pearson correlation coefficient p < 0.001) between both methods, although the absolute concentration determined is different from factor 5. The validation of our SRM method encourages us to propose it as an alternative approach for accurate determination of hepcidin in human samples for clinical diagnosis, follow-up and management of iron-related diseases.
topic Hepcidin
Iron deficiency
Mass spectrometry
ELISA
url http://www.sciencedirect.com/science/article/pii/S2212968514000087
work_keys_str_mv AT constancedelaby clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT jeromevialaret clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT paulinebros clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT audreygabelle clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT thibaudlefebvre clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT hervepuy clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT christophehirtz clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
AT sylvainlehmann clinicalmeasurementofhepcidin25inhumanserumisquantitativemassspectrometryuptothejob
_version_ 1716782780718776320