A novel immunological assay for hepcidin quantification in human serum.
BACKGROUND: Hepcidin is a 25-aminoacid cysteine-rich iron regulating peptide. Increased hepcidin concentrations lead to iron sequestration in macrophages, contributing to the pathogenesis of anaemia of chronic disease whereas decreased hepcidin is observed in iron deficiency and primary iron overloa...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2009-01-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC2640459?pdf=render |
id |
doaj-0a484668243c496b921ee0309b75c366 |
---|---|
record_format |
Article |
spelling |
doaj-0a484668243c496b921ee0309b75c3662020-11-25T02:28:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-01-0142e458110.1371/journal.pone.0004581A novel immunological assay for hepcidin quantification in human serum.Vasiliki KoliarakiMartha MarinouTheodoros P VassilakopoulosEustathios VavourakisEmmanuel TsochatzisGerassimos A PangalisGeorge PapatheodoridisAlexandra StamoulakatouDorine W SwinkelsGeorge PapanikolaouAvgi MamalakiBACKGROUND: Hepcidin is a 25-aminoacid cysteine-rich iron regulating peptide. Increased hepcidin concentrations lead to iron sequestration in macrophages, contributing to the pathogenesis of anaemia of chronic disease whereas decreased hepcidin is observed in iron deficiency and primary iron overload diseases such as hereditary hemochromatosis. Hepcidin quantification in human blood or urine 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 a specific and non-operator demanding immunoassay for hepcidin quantification in human sera. METHODS AND FINDINGS: An ELISA assay was developed for measuring hepcidin serum concentration using a recombinant hepcidin25-His peptide and a polyclonal antibody against this peptide, which was able to identify native hepcidin. The ELISA assay had a detection range of 10-1500 microg/L and a detection limit of 5.4 microg/L. The intra- and interassay coefficients of variance ranged from 8-15% and 5-16%, respectively. Mean linearity and recovery were 101% and 107%, respectively. Mean hepcidin levels were significantly lower in 7 patients with juvenile hemochromatosis (12.8 microg/L) and 10 patients with iron deficiency anemia (15.7 microg/L) and higher in 7 patients with Hodgkin lymphoma (116.7 microg/L) compared to 32 age-matched healthy controls (42.7 microg/L). CONCLUSIONS: We describe a new simple ELISA assay for measuring hepcidin in human serum with sufficient accuracy and reproducibility.http://europepmc.org/articles/PMC2640459?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Vasiliki Koliaraki Martha Marinou Theodoros P Vassilakopoulos Eustathios Vavourakis Emmanuel Tsochatzis Gerassimos A Pangalis George Papatheodoridis Alexandra Stamoulakatou Dorine W Swinkels George Papanikolaou Avgi Mamalaki |
spellingShingle |
Vasiliki Koliaraki Martha Marinou Theodoros P Vassilakopoulos Eustathios Vavourakis Emmanuel Tsochatzis Gerassimos A Pangalis George Papatheodoridis Alexandra Stamoulakatou Dorine W Swinkels George Papanikolaou Avgi Mamalaki A novel immunological assay for hepcidin quantification in human serum. PLoS ONE |
author_facet |
Vasiliki Koliaraki Martha Marinou Theodoros P Vassilakopoulos Eustathios Vavourakis Emmanuel Tsochatzis Gerassimos A Pangalis George Papatheodoridis Alexandra Stamoulakatou Dorine W Swinkels George Papanikolaou Avgi Mamalaki |
author_sort |
Vasiliki Koliaraki |
title |
A novel immunological assay for hepcidin quantification in human serum. |
title_short |
A novel immunological assay for hepcidin quantification in human serum. |
title_full |
A novel immunological assay for hepcidin quantification in human serum. |
title_fullStr |
A novel immunological assay for hepcidin quantification in human serum. |
title_full_unstemmed |
A novel immunological assay for hepcidin quantification in human serum. |
title_sort |
novel immunological assay for hepcidin quantification in human serum. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2009-01-01 |
description |
BACKGROUND: Hepcidin is a 25-aminoacid cysteine-rich iron regulating peptide. Increased hepcidin concentrations lead to iron sequestration in macrophages, contributing to the pathogenesis of anaemia of chronic disease whereas decreased hepcidin is observed in iron deficiency and primary iron overload diseases such as hereditary hemochromatosis. Hepcidin quantification in human blood or urine 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 a specific and non-operator demanding immunoassay for hepcidin quantification in human sera. METHODS AND FINDINGS: An ELISA assay was developed for measuring hepcidin serum concentration using a recombinant hepcidin25-His peptide and a polyclonal antibody against this peptide, which was able to identify native hepcidin. The ELISA assay had a detection range of 10-1500 microg/L and a detection limit of 5.4 microg/L. The intra- and interassay coefficients of variance ranged from 8-15% and 5-16%, respectively. Mean linearity and recovery were 101% and 107%, respectively. Mean hepcidin levels were significantly lower in 7 patients with juvenile hemochromatosis (12.8 microg/L) and 10 patients with iron deficiency anemia (15.7 microg/L) and higher in 7 patients with Hodgkin lymphoma (116.7 microg/L) compared to 32 age-matched healthy controls (42.7 microg/L). CONCLUSIONS: We describe a new simple ELISA assay for measuring hepcidin in human serum with sufficient accuracy and reproducibility. |
url |
http://europepmc.org/articles/PMC2640459?pdf=render |
work_keys_str_mv |
AT vasilikikoliaraki anovelimmunologicalassayforhepcidinquantificationinhumanserum AT marthamarinou anovelimmunologicalassayforhepcidinquantificationinhumanserum AT theodorospvassilakopoulos anovelimmunologicalassayforhepcidinquantificationinhumanserum AT eustathiosvavourakis anovelimmunologicalassayforhepcidinquantificationinhumanserum AT emmanueltsochatzis anovelimmunologicalassayforhepcidinquantificationinhumanserum AT gerassimosapangalis anovelimmunologicalassayforhepcidinquantificationinhumanserum AT georgepapatheodoridis anovelimmunologicalassayforhepcidinquantificationinhumanserum AT alexandrastamoulakatou anovelimmunologicalassayforhepcidinquantificationinhumanserum AT dorinewswinkels anovelimmunologicalassayforhepcidinquantificationinhumanserum AT georgepapanikolaou anovelimmunologicalassayforhepcidinquantificationinhumanserum AT avgimamalaki anovelimmunologicalassayforhepcidinquantificationinhumanserum AT vasilikikoliaraki novelimmunologicalassayforhepcidinquantificationinhumanserum AT marthamarinou novelimmunologicalassayforhepcidinquantificationinhumanserum AT theodorospvassilakopoulos novelimmunologicalassayforhepcidinquantificationinhumanserum AT eustathiosvavourakis novelimmunologicalassayforhepcidinquantificationinhumanserum AT emmanueltsochatzis novelimmunologicalassayforhepcidinquantificationinhumanserum AT gerassimosapangalis novelimmunologicalassayforhepcidinquantificationinhumanserum AT georgepapatheodoridis novelimmunologicalassayforhepcidinquantificationinhumanserum AT alexandrastamoulakatou novelimmunologicalassayforhepcidinquantificationinhumanserum AT dorinewswinkels novelimmunologicalassayforhepcidinquantificationinhumanserum AT georgepapanikolaou novelimmunologicalassayforhepcidinquantificationinhumanserum AT avgimamalaki novelimmunologicalassayforhepcidinquantificationinhumanserum |
_version_ |
1724838000440901632 |