Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin

Nicole U. Stoffel; Christophe Zeder; Eloïse Fort; Dorine W. Swinkels; Michael B. Zimmermann; Diego Moretti

doi:10.1515/cclm-2017-0097

Artikel

Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin

Nicole U. Stoffel , Christophe Zeder , Eloïse Fort , Dorine W. Swinkels , Michael B. Zimmermann und Diego Moretti

Veröffentlicht/Copyright: 17. Juni 2017

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Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 55 Heft 8

Abstract

Background:

Hepcidin is the central systemic regulator of iron metabolism, but its quantification in biological fluids is challenging. Rapid, accurate and user-friendly methods are needed. Our aim was to assess the ability of hepcidin as measured by three different c-ELISA assays to predict iron bioavailability in humans.

Methods:

The three assays used were commercially available DRG and Peninsula assays and the c-ELISA method performed at Radboud University Medical Centre, Nijmegen, The Netherlands (Hepcidinanalysis.com), validated by comparative measurements with time-of-flight mass spectrometry. We analyzed plasma samples (n=37) selected to represent a broad range of hepcidin concentrations from a subgroup of healthy, iron-depleted women in a study assessing fractional absorption from iron supplements.

Results:

In single regressions, all three c-ELISA assays were predictors of fractional iron absorption: R²=0.363 (DRG), R²=0.281 (Peninsula) and R²=0.327 (Hepcidinanalysis.com). In multiple regressions, models including hepcidin measured with either DRG-, Peninsula or Hepcidinanalysis.com explained 55.7%, 44.5% and 52.5% of variance in fractional absorption, and hepcidin was a strong predictor of fractional absorption irrespective of the hepcidin assays used. However, we found significant differences in absolute values for hepcidin between different methods. Both the DRG assay’s (y=0.61x+0.87; R²=0.873) and the Peninsula assay’s measurements (y=1.88x+0.62; R²=0.770) were correlated with Hepcidinanalysis.com.

Conclusions:

The biological variability in plasma hepcidin, (inter-sample CV) was 5–10-fold higher for both the Peninsula and DRG assay than the analytical variably (inter-run within-sample CV) suggesting substantial discriminatory power to distinguish biological hepcidin variation. Between methods, prediction of iron bioavailability in generally healthy iron depleted subjects appears comparable.

Keywords: anemia of chronic disease; hepcidin; iron absorption; iron deficiency anemia; iron overload

Acknowledgments

We would like to thank Andrew Armitage, Weatherall Institute of Molecular Medicine University of Oxford, Oxford, UK for very valuable analytical advice on the calibration of the Peninsula measurement kit.

Author contributions: NUS and DM designed the study. NUS and EF did the hepcidin measurements with the two commercially available c-ELISA assays. NUS analyzed the data and wrote the first draft of the manuscript. All authors contributed to the final version of the manuscript. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: DWS is an employee of Radboud University and Medical Centre that offers high quality hepcidin measurements to the scientific, medical and pharmaceutical community, at a fee for service basis.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2017-2-1

Accepted: 2017-4-25

Published Online: 2017-6-17

Published in Print: 2017-7-26

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Artikel in diesem Heft

https://doi.org/10.1515/cclm-2017-0097

Schlagwörter für diesen Artikel

anemia of chronic disease; hepcidin; iron absorption; iron deficiency anemia; iron overload