Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.

Erythroferrone (ERFE), the erythroid regulator of iron metabolism, inhibits hepcidin to increase iron availability for erythropoiesis. ERFE plays a pathological role during ineffective erythropoiesis as occurs in X-linked sideroblastic anemia (XLSA) and β-thalassemia. Its measurement might serve as...

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Autores principales:	Laura Diepeveen, Rian Roelofs, Nicolai Grebenchtchikov, Rachel van Swelm, Leon Kautz, Dorine Swinkels
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/173e5fa918e9448cb8c2232858c56c27
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spelling	oai:doaj.org-article:173e5fa918e9448cb8c2232858c56c272021-12-02T20:06:47ZDifferentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.1932-620310.1371/journal.pone.0254851https://doaj.org/article/173e5fa918e9448cb8c2232858c56c272021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0254851https://doaj.org/toc/1932-6203Erythroferrone (ERFE), the erythroid regulator of iron metabolism, inhibits hepcidin to increase iron availability for erythropoiesis. ERFE plays a pathological role during ineffective erythropoiesis as occurs in X-linked sideroblastic anemia (XLSA) and β-thalassemia. Its measurement might serve as an indicator of severity for these diseases. However, for reliable quantification of ERFE analytical characterization is indispensable to determine the assay's limitations and define proper methodology. We developed a sandwich ELISA for human serum ERFE using polyclonal antibodies and report its extensive analytical validation. This new assay showed, for the first time, the differentiation of XLSA and β-thalassemia major patients from healthy controls (p = 0.03) and from each other (p<0.01), showing the assay provides biological plausible results. Despite poor dilution linearity, parallelism and recovery in patient serum matrix, which indicated presence of a matrix effect and/or different immunoreactivity of the antibodies to the recombinant standard and the endogenous analyte, our assay correlated well with two other existing ERFE ELISAs (both R2 = 0.83). Nevertheless, employment of one optimal dilution of all serum samples is warranted to obtain reliable results. When adequately performed, the assay can be used to further unravel the human erythropoiesis-hepcidin-iron axis in various disorders and assess the added diagnostic value of ERFE.Laura DiepeveenRian RoelofsNicolai GrebenchtchikovRachel van SwelmLeon KautzDorine SwinkelsPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 7, p e0254851 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Laura Diepeveen Rian Roelofs Nicolai Grebenchtchikov Rachel van Swelm Leon Kautz Dorine Swinkels Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.
description	Erythroferrone (ERFE), the erythroid regulator of iron metabolism, inhibits hepcidin to increase iron availability for erythropoiesis. ERFE plays a pathological role during ineffective erythropoiesis as occurs in X-linked sideroblastic anemia (XLSA) and β-thalassemia. Its measurement might serve as an indicator of severity for these diseases. However, for reliable quantification of ERFE analytical characterization is indispensable to determine the assay's limitations and define proper methodology. We developed a sandwich ELISA for human serum ERFE using polyclonal antibodies and report its extensive analytical validation. This new assay showed, for the first time, the differentiation of XLSA and β-thalassemia major patients from healthy controls (p = 0.03) and from each other (p<0.01), showing the assay provides biological plausible results. Despite poor dilution linearity, parallelism and recovery in patient serum matrix, which indicated presence of a matrix effect and/or different immunoreactivity of the antibodies to the recombinant standard and the endogenous analyte, our assay correlated well with two other existing ERFE ELISAs (both R2 = 0.83). Nevertheless, employment of one optimal dilution of all serum samples is warranted to obtain reliable results. When adequately performed, the assay can be used to further unravel the human erythropoiesis-hepcidin-iron axis in various disorders and assess the added diagnostic value of ERFE.
format	article
author	Laura Diepeveen Rian Roelofs Nicolai Grebenchtchikov Rachel van Swelm Leon Kautz Dorine Swinkels
author_facet	Laura Diepeveen Rian Roelofs Nicolai Grebenchtchikov Rachel van Swelm Leon Kautz Dorine Swinkels
author_sort	Laura Diepeveen
title	Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.
title_short	Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.
title_full	Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.
title_fullStr	Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.
title_full_unstemmed	Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.
title_sort	differentiating iron-loading anemias using a newly developed and analytically validated elisa for human serum erythroferrone.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/173e5fa918e9448cb8c2232858c56c27
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Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone.

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