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Background: Hepcidin is a 25-aminoacid cysteine-rich iron regulating peptide. Hepcidin quantification in human serum provides new insights for the pathogenesis of disorders of iron homeostasis. This study describes an ELISA immunoassay for hepcidin quantification in human serum and reference ranges for Bulgarian population.
Methods: We used a sandwich ELISA method from USCN Life Science inc. that consists of ready to use, precoated 96-well strip plate with 2 antihepcidin-25 monoclonal antibodies. A recombinant Hepcidin in 16 μg/L concentration is used as a standard. We correlated ELISA results of hepcidin-25 measurements in healthy population to ferritin, hemoglobin concentration in reticulocytes, transferrin, and iron levels.
Results: The sandwich ELISA was highly specific for hepcidin-25. We found that serum hepcidin levels for Bulgarian population are 3.052 µg/L - 37.750 µg/L, which is quite similar to that established by WCX-TOF MS from the Laboratory of Genetic, Endocrine and Metabolic Diseases; Dept. of Laboratory Medicine, Radbound University Medical Centre; Nijmegen, The Netherlands. Ferritin levels and hemoglobin concentration in reticulocytes correlated significantly to serum hepcidin levels (0.3 < r < 0.5, p < 0.010). Transferrin levels showed negative and no significant correlation to hepcidin in serum (r = -0.111).
Conclusions: The use of two monoclonal antibodies in a sandwich ELISA format provides a reliable, reproducible, and not very expensive method for measuring serum concentrations of the bioactive form of hepcidin in Bulgarian laboratory practice.
DOI: 10.7754/Clin.Lab.2014.140322
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