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

Stoffel, Nicole U. and Zeder, Christophe and Fort, Eloise and Swinkels, Dorine W. and Zimmermann, Michael B. and Moretti, Diego (2017) Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin. CLINICAL CHEMISTRY AND LABORATORY MEDICINE, 55 (8). pp. 1186-1192.

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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: R-2 = 0.363 (DRG), R-2 = 0.281 (Peninsula) and R-2 = 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; R-2 = 0.873) and the Peninsula assay's measurements (y = 1.88x + 0.62; R-2 = 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.

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