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Acta Medica Bulgarica

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0324-1750
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Elisa Method For Serum Hepcidin Quantification In Bulgarian Population

V. Manolov
  • Department of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, Bulgaria
  • Other articles by this author:
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/ B. Atanasova
  • Department of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, Bulgaria
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ V. Vasilev
  • Department of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, Bulgaria
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ K. Tzatchev
  • Department of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, Bulgaria
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  • De Gruyter OnlineGoogle Scholar
/ M. Velizarova
  • Department of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, Bulgaria
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2014-11-05 | DOI: https://doi.org/10.2478/amb-2014-0003

Summary

Hepcidin is a 25-aminoacid cysteine-rich iron regulating peptide. Hepcidin quantification in human blood may provide further insights for the pathogenesis of disorders of iron homeostasis and might prove a valuable tool for clinicians for the differential diagnosis of anaemia. This study describes ELISA immunoassay for hepcidin quantification in human serum. We used a sandwich ELISA method from USCN Life Science inc., that consists of ready to use, pre-coated 96-well strip plate with 2 antihepcidin-25 monoclonal antibodies. A recombinant hepcidin in 16 μg/l concentration is used as a standard; it reconstitutes with 1.0 ml standard diluent to prepare a stock solution. We correlated ELISA results of hepcidin-25 measurements in healthy population with hemodialysis patients. The sandwich ELISA was highly specific for hepcidin-25, having a low limit of quantification of 0.020 μg/l. Hepcidin- 25 concentrations were increased in hemodialysis patients (median 33.05 μg/l, range 22.31 -60.98 μg/l, n = 10) compared with healthy individuals (median 12.41 μg/l, range 6.05-18.53 μg/l, n = 40). The use of 2 monoclonal antibodies in a sandwich ELISA format provides a robust, convenient and not very expensive method for measuring concentrations of the active form of hepcidin. It should help to improve our understanding of the role of hepcidin in regulating iron metabolism.

Keywords : hepcidin; quantification; ELISA; hemodialysis; iron metabolism

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About the article

Published Online: 2014-11-05

Published in Print: 2014-06-01


Citation Information: Acta Medica Bulgarica, ISSN (Online) 0324-1750, DOI: https://doi.org/10.2478/amb-2014-0003.

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© 2014. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

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