A dual-monoclonal sandwich ELISA specific for hepcidin-25
Abstract
BACKGROUND: Hepcidin, a key regulator of iron metabolism, binds to the iron transporter ferroportin to cause its degradation. In humans, hepcidin deficiency has been linked to hemochromatosis and iron overload, whereas increased concentrations have been reported in anemia of cancer and chronic disease. There is currently an unmet clinical need for a specific immunoassay with a low limit of quantification to measure serum concentrations of hepcidin-25, the active form of the protein. METHODS: We generated 2 antihepcidin-25 monoclonal antibodies and used them to build a sandwich ELISA. We correlated ELISA results to hepcidin-25 measurements by LC-MS and used ELISA to measure serum hepcidin-25 concentrations in normal individuals, cancer patients, and patients with rheumatoid arthritis. RESULTS: The sandwich ELISA was highly specific for hepcidin-25, having a limit of quantification of 0.01 μg/L (10 pg/mL). Serum concentrations of hepcidin-25 measured by ELISA correlated with hepcidin-25 concentrations measured by using an independent LC-MS assay (r = 0.98, P < 0.001). Hepcidin-25 concentrations were increased in patients with cancer (median 54.8 μg/L, 25%-75% range 23.2-93.5 μg/L, n = 34) and rheumatoid arthritis (median 10.6 μg/L, 25%-75% range 5.9-18.4 μg/L, n = 76) compared with healthy individuals (median 1.20 μg/L, 25%-75% range 0.42-3.07 μg/L, n = 100). CONCLUSIONS: The use of 2 monoclonal antibodies in a sandwich ELISA format provides a robust and convenient method for measuring concentrations of the active form of hepcidin. This ELISA should help to improve our understanding of the role of hepcidin in regulating iron metabolism.
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In cancer patients, hepcidin levels are often elevated, likely because of cancer driven inflammation.
“In cancer patients, hepcidin levels are often elevated, likely because of cancer driven inflammation [48,49].”
From Iron as a central player and promising target in cancer progression (2019) · cites this paper as [49]
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Słomka, A., Koba, M., Kulwas, A., & Żekanowska, E. (2011) · Current Pharmaceutical Analysis · doi:10.2174/157341211796353309 · Cited by 2 · see citing works
Kawabata, H., Miyazawa, N., Matsuda, Y., Satobe, M., Mizogami, Y., Kuriyama, Y., Sakai, T., Mori, M., & Sasa, M. (2024) · International Journal of Hematology · doi:10.1007/s12185-024-03720-4 · Cited by 2 · see citing works
Zhou, Y., Li, T., Zhu, L., Lu, Z., Peng, W., Hu, Y., Liu, J., Yuan, J., & Zhu, Y. (2026) · Journal of Trace Elements in Medicine and Biology · doi:10.1016/j.jtemb.2026.127875 · Cited by 1 · see citing works
Laboratory Assessment of Iron Status
Brugnara, C. (2019) · Cambridge University Press eBooks · doi:10.1017/9781139023993.006
Podgórski, P., Przybyłkowski, A. M., Nehring, P., & Ciepiela, O. (2025) · Digestive Diseases · doi:10.1159/000549784
- Authors
- Butterfield, A. M., Luan, P., Witcher, D. R., Manetta, J., Murphy, A. T., Wroblewski, V. J., & Konrad, R. J.
- Affiliations
- Eli Lilly (United States) (US)
- Corresponding author
- Konrad, Robert J
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2010-11-01
- Volume
- 56
- Issue
- 11
- Pages
- 1725-1732
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W2122332989
- Cited by
- 59
- References
- 37
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 1 · 2021: 3 · 2020: 2 · 2019: 3 · 2018: 4 · 2017: 3 · 2016: 5 · 2015: 5 · 2014: 9 · 2013: 10 · 2012: 5
- FWCI
- 4.00
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Clinical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Ferroptosis and cancer prognosis
- Health Sciences › Medicine › Hematology › Erythropoietin and Anemia Treatment
- Keywords
- hepcidin-25, sandwich ELISA, monoclonal antibodies, immunoassay development, iron metabolism, cancer, rheumatoid arthritis, anemia of chronic disease, ferroportin, hemochromatosis
- MeSH terms
- Adolescent, Adult, Antibodies, Monoclonal, Arthritis, Rheumatoid, Enzyme-Linked Immunosorbent Assay, Female, Humans, Male, Middle Aged, Neoplasms, Reference Values, Sex Factors, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Antimicrobial Cationic Peptides, Racial Groups, Young Adult, Hepcidins
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