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

AbstractFrom 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, 2ng/mL), limit of quantification (LOQ, 6ng/mL), repeatability, reproducibility and linearity (up to 200ng/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