Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.

<h4>Background</h4>Bioavailable 25-hydroxyvitamin D (25OHD) may be a better indicator of vitamin D sufficiency than total 25OHD. This report describes a novel assay for measuring serum bioavailable 25OHD.<h4>Methods</h4>We developed an assay for 25OHD % bioavailability based...

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Autores principales:	Anders H Berg, Mahtab Tavasoli, Agnes S Lo, Sherri-Ann M Burnett-Bowie, Ishir Bhan, S Ananth Karumanchi, Sahir Kalim, Dongsheng Zhang, Sophia Zhao, Ravi I Thadhani
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6090c91ee93c4db39ab2de7fba8ece91
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spelling	oai:doaj.org-article:6090c91ee93c4db39ab2de7fba8ece912021-12-02T20:09:21ZDevelopment and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.1932-620310.1371/journal.pone.0254158https://doaj.org/article/6090c91ee93c4db39ab2de7fba8ece912021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0254158https://doaj.org/toc/1932-6203<h4>Background</h4>Bioavailable 25-hydroxyvitamin D (25OHD) may be a better indicator of vitamin D sufficiency than total 25OHD. This report describes a novel assay for measuring serum bioavailable 25OHD.<h4>Methods</h4>We developed an assay for 25OHD % bioavailability based on competitive binding of 25OHD tracer between vitamin D-binding protein (DBP)-coated affinity chromatography beads and serum DBP. Bioavailable 25OHD, total 25OHD, albumin, and DBP protein concentrations were measured in 89 samples from hospitalized patients and 42 healthy controls to determine how the DBP binding assay responds to differences in concentrations of DBP and compares to calculated bioavailable 25OHD values.<h4>Results</h4>DBP binding assay showed a linear relationship between DBP-bound 25OHD tracer recovered from bead supernatant and DBP calibrator concentrations (y = 0.0017x +0.731, R2 = 0.9961, p<0.001). Inversion of this relationship allowed interpolation of DBP binding equivalents based upon 25OHD tracer recovered. The relationship between DBP binding equivalents and % bioavailability fits a non-linear curve, allowing calculation of % bioavailable 25OHD from DBP binding equivalents (y = 10.625x-0.817, R2 = 0.9961, p<0.001). In hospitalized patient samples, there were linear relationships between DBP protein concentrations and DBP binding equivalents (y = 0.7905x + 59.82, R2 = 0.8597, p<0.001), between measured vs. calculated % bioavailability (y = 0.9528 + 0.0357, R2 = 0.7200, p<0.001), and between absolute concentrations of measured vs. calculated bioavailable 25OHD (y = 1.2403 + 0.1221, R2 = 0.8913, p<0.001).<h4>Conclusions</h4>The DBP-binding assay for bioavailable 25OHD shows expected changes in 25OHD % bioavailability in response to changes in DBP concentrations and concordance with calculated bioavailable 25OHD concentrations.Anders H BergMahtab TavasoliAgnes S LoSherri-Ann M Burnett-BowieIshir BhanS Ananth KarumanchiSahir KalimDongsheng ZhangSophia ZhaoRavi I ThadhaniPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 7, p e0254158 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Anders H Berg Mahtab Tavasoli Agnes S Lo Sherri-Ann M Burnett-Bowie Ishir Bhan S Ananth Karumanchi Sahir Kalim Dongsheng Zhang Sophia Zhao Ravi I Thadhani Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.
description	<h4>Background</h4>Bioavailable 25-hydroxyvitamin D (25OHD) may be a better indicator of vitamin D sufficiency than total 25OHD. This report describes a novel assay for measuring serum bioavailable 25OHD.<h4>Methods</h4>We developed an assay for 25OHD % bioavailability based on competitive binding of 25OHD tracer between vitamin D-binding protein (DBP)-coated affinity chromatography beads and serum DBP. Bioavailable 25OHD, total 25OHD, albumin, and DBP protein concentrations were measured in 89 samples from hospitalized patients and 42 healthy controls to determine how the DBP binding assay responds to differences in concentrations of DBP and compares to calculated bioavailable 25OHD values.<h4>Results</h4>DBP binding assay showed a linear relationship between DBP-bound 25OHD tracer recovered from bead supernatant and DBP calibrator concentrations (y = 0.0017x +0.731, R2 = 0.9961, p<0.001). Inversion of this relationship allowed interpolation of DBP binding equivalents based upon 25OHD tracer recovered. The relationship between DBP binding equivalents and % bioavailability fits a non-linear curve, allowing calculation of % bioavailable 25OHD from DBP binding equivalents (y = 10.625x-0.817, R2 = 0.9961, p<0.001). In hospitalized patient samples, there were linear relationships between DBP protein concentrations and DBP binding equivalents (y = 0.7905x + 59.82, R2 = 0.8597, p<0.001), between measured vs. calculated % bioavailability (y = 0.9528 + 0.0357, R2 = 0.7200, p<0.001), and between absolute concentrations of measured vs. calculated bioavailable 25OHD (y = 1.2403 + 0.1221, R2 = 0.8913, p<0.001).<h4>Conclusions</h4>The DBP-binding assay for bioavailable 25OHD shows expected changes in 25OHD % bioavailability in response to changes in DBP concentrations and concordance with calculated bioavailable 25OHD concentrations.
format	article
author	Anders H Berg Mahtab Tavasoli Agnes S Lo Sherri-Ann M Burnett-Bowie Ishir Bhan S Ananth Karumanchi Sahir Kalim Dongsheng Zhang Sophia Zhao Ravi I Thadhani
author_facet	Anders H Berg Mahtab Tavasoli Agnes S Lo Sherri-Ann M Burnett-Bowie Ishir Bhan S Ananth Karumanchi Sahir Kalim Dongsheng Zhang Sophia Zhao Ravi I Thadhani
author_sort	Anders H Berg
title	Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.
title_short	Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.
title_full	Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.
title_fullStr	Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.
title_full_unstemmed	Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.
title_sort	development and analytical validation of a novel bioavailable 25-hydroxyvitamin d assay.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/6090c91ee93c4db39ab2de7fba8ece91
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Development and analytical validation of a novel bioavailable 25-hydroxyvitamin D assay.

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