Verification of the Automated ELISA Assay for Hepcidin-25 in Human Serum

Abstract

Introduction: Hepcidin-25, the bioactive form of hepcidin, is the master protein in regulating iron homeostasis. Serum concentrations, measured by different methods, are often incomparable and complicate results interpretation. Materials and Methods: The aim was to verify the first fully automated enzyme-linked immunosorbent assay (ELISA) method, using the DRG Hybrid XL analyzer (DRG Instruments, Marburg, Germany) standardized against the mass spectrometry method. Intra- (CVi) and inter-assay (CVg) precision and bias were performed using commercially available controls with low (C1) and high (C2) concentrations. The reference interval was verified by analyzing serum samples of 20 healthy males. Results: CVi = 9.1% (C1), 4.5% (C2); CVg = 8.9% (C1), 5.6% (C2); calculated bias was 33% for C1 and 20% for C2, respectively. Conclusion: Verification of the fully automated ELISA method for hepcidin-25 in serum on the DRG Hybrid XL analyzer met the analytical acceptance criteria.

Keywords

• enzyme-linked immunosorbent assay
• hepcidins
• homeostasis
• iron