caa a22 4500 445888946 CHVBK 20180317145609.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s00535-010-0344-z doi (NATIONALLICENCE)springer-10.1007/s00535-010-0344-z A novel monoclonal antibody immunoassay for the detection of human serum hepcidin [Elektronische Daten] [Peggy Schwarz, Pavel Strnad, Guido von Figura, Alfred Janetzko, Pierre Krayenbühl, Guido Adler, Hasan Kulaksiz] Background: Hepcidin is a liver-derived peptide hormone regulating iron metabolism. Changes in the expression of hepcidin are known to be the key pathogenic factors in hereditary hemochromatosis and are associated with infection and inflammation. To better understand the hormone's function in human disease, we aimed to establish an immunoassay to determine hepcidin concentrations in serum. Methods: Monoclonal antibodies mHK(8) and mHK(9) were generated and characterized by dot blot, Western blot, and immunofluorescence. A competitive enzyme-linked immunosorbent assay (ELISA) was established with mHK(9). Results: Both antibodies recognized hepcidin, by dot blot and Western blot, respectively. In human liver, mHK(8)/(9) showed an immunofluorescence staining pattern in hepatocytes identical to that of established prohepcidin antibodies. The developed immunoassay with mHK(9), reliably detecting mature hepcidin in serum over a large concentration range (0.9-140ngml−1), showed high sensitivity and precision (intra-/interassay coefficients of variation: 4-5 and 7-11%; mean linearity: 85-112%; mean recovery: 87-114%). To test the clinical functionality of the developed assay we measured hepcidin serum concentrations in healthy volunteers, hepatitis C virus (HCV) patients, and two groups of hemochromatotic patients undergoing phlebotomy. The assay distinguished low hepcidin level in HCV and homozygous hemochromatosis patients from normal-range controls and compound heterozygous hemochromatosis patients. In healthy subjects and HCV patients, hepcidin levels were correlated with iron and transferrin saturation; no correlation was observed in the hemochromatotic patients. Conclusion: We developed a monoclonal antibody ELISA that quantifies serum hepcidin levels with high sensitivity, robustness, and reliability of detection. The hepcidin ELISA should help to enhance our understanding of hepcidin-related human disorders. Springer, 2010 Hepcidin nationallicence ELISA nationallicence Hemochromatosis nationallicence Iron nationallicence Liver nationallicence Schwarz Peggy Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut Strnad Pavel Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut von Figura Guido Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut Janetzko Alfred DRG Instruments GmbH, Marburg, Germany aut Krayenbühl Pierre Department of Internal Medicine, University Hospital, Zurich, Switzerland aut Adler Guido Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut Kulaksiz Hasan Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut Journal of Gastroenterology Springer Japan 46/5(2011-05-01), 648-656 0944-1174 46:5<648 2011 46 535 https://doi.org/10.1007/s00535-010-0344-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00535-010-0344-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Schwarz Peggy Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut NATIONALLICENCE 700 1- Strnad Pavel Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut NATIONALLICENCE 700 1- von Figura Guido Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut NATIONALLICENCE 700 1- Janetzko Alfred DRG Instruments GmbH, Marburg, Germany aut NATIONALLICENCE 700 1- Krayenbühl Pierre Department of Internal Medicine, University Hospital, Zurich, Switzerland aut NATIONALLICENCE 700 1- Adler Guido Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut NATIONALLICENCE 700 1- Kulaksiz Hasan Division of Gastroenterology, Department of Internal Medicine, University Hospital Ulm, Albert-Einstein-Allee 23, 89081, Ulm, Germany aut NATIONALLICENCE 773 0- Journal of Gastroenterology Springer Japan 46/5(2011-05-01), 648-656 0944-1174 46:5<648 2011 46 535 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer