Elisa Method For Serum Hepcidin Quantification In Bulgarian Population
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 E...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Sciendo
2014-06-01
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| Series: | Acta Medica Bulgarica |
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| Online Access: | https://doi.org/10.2478/amb-2014-0003 |
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| author | Manolov V. Atanasova B. Vasilev V. Tzatchev K. Velizarova M. |
| author_facet | Manolov V. Atanasova B. Vasilev V. Tzatchev K. Velizarova M. |
| author_sort | Manolov V. |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T10:40:27Z |
| format | Article |
| id | doaj.art-dbc2722631b04b03a6e4a28f688103be |
| institution | Directory Open Access Journal |
| issn | 0324-1750 |
| language | English |
| last_indexed | 2024-12-17T10:40:27Z |
| publishDate | 2014-06-01 |
| publisher | Sciendo |
| record_format | Article |
| series | Acta Medica Bulgarica |
| spelling | doaj.art-dbc2722631b04b03a6e4a28f688103be2022-12-21T21:52:16ZengSciendoActa Medica Bulgarica0324-17502014-06-01411222910.2478/amb-2014-0003amb-2014-0003Elisa Method For Serum Hepcidin Quantification In Bulgarian PopulationManolov V.0Atanasova B.1Vasilev V.2Tzatchev K.3Velizarova M.4Department of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, BulgariaDepartment of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, BulgariaDepartment of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, BulgariaDepartment of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, BulgariaDepartment of Clinical Laboratory and Clinical Immunology, Medical Faculty, Medical University, Sofia, BulgariaHepcidin 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.https://doi.org/10.2478/amb-2014-0003hepcidinquantificationelisahemodialysisiron metabolism |
| spellingShingle | Manolov V. Atanasova B. Vasilev V. Tzatchev K. Velizarova M. Elisa Method For Serum Hepcidin Quantification In Bulgarian Population Acta Medica Bulgarica hepcidin quantification elisa hemodialysis iron metabolism |
| title | Elisa Method For Serum Hepcidin Quantification In Bulgarian Population |
| title_full | Elisa Method For Serum Hepcidin Quantification In Bulgarian Population |
| title_fullStr | Elisa Method For Serum Hepcidin Quantification In Bulgarian Population |
| title_full_unstemmed | Elisa Method For Serum Hepcidin Quantification In Bulgarian Population |
| title_short | Elisa Method For Serum Hepcidin Quantification In Bulgarian Population |
| title_sort | elisa method for serum hepcidin quantification in bulgarian population |
| topic | hepcidin quantification elisa hemodialysis iron metabolism |
| url | https://doi.org/10.2478/amb-2014-0003 |
| work_keys_str_mv | AT manolovv elisamethodforserumhepcidinquantificationinbulgarianpopulation AT atanasovab elisamethodforserumhepcidinquantificationinbulgarianpopulation AT vasilevv elisamethodforserumhepcidinquantificationinbulgarianpopulation AT tzatchevk elisamethodforserumhepcidinquantificationinbulgarianpopulation AT velizarovam elisamethodforserumhepcidinquantificationinbulgarianpopulation |