A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum

Abstract Calciprotein particles, nanoscale aggregates of insoluble mineral and binding proteins, have emerged as potential mediators of phosphate toxicity in patients with Chronic Kidney Disease. Although existing immunochemical methods for their detection have provided compelling data, these approa...

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Autores principales:	Edward R. Smith, Tim D. Hewitson, Michael M. X. Cai, Parisa Aghagolzadeh, Matthias Bachtler, Andreas Pasch, Stephen G. Holt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4660db06f2274211a15dd40111002799
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spelling	oai:doaj.org-article:4660db06f2274211a15dd401110027992021-12-02T12:32:50ZA novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum10.1038/s41598-017-05474-y2045-2322https://doaj.org/article/4660db06f2274211a15dd401110027992017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05474-yhttps://doaj.org/toc/2045-2322Abstract Calciprotein particles, nanoscale aggregates of insoluble mineral and binding proteins, have emerged as potential mediators of phosphate toxicity in patients with Chronic Kidney Disease. Although existing immunochemical methods for their detection have provided compelling data, these approaches are indirect, lack specificity and are subject to a number of other technical and theoretical shortcomings. Here we have developed a rapid homogeneous fluorescent probe-based flow cytometric method for the detection and quantitation of individual mineral-containing nanoparticles in human and animal serum. This method allows the discrimination of membrane-bound from membrane-free particles and different mineral phases (amorphous vs. crystalline). Critically, the method has been optimised for use on a conventional instrument, without the need for manual hardware adjustments. Using this method, we demonstrate a consistency in findings across studies of Chronic Kidney Disease patients and commonly used uraemic animal models. These studies demonstrate that renal dysfunction is associated with the ripening of calciprotein particles to the crystalline state and reveal bone metabolism and dietary mineral as important modulators of circulating levels. Flow cytometric analysis of calciprotein particles may enhance our understanding of mineral handling in kidney disease and provide a novel indicator of therapeutic efficacy for interventions targeting Chronic Kidney Disease-Mineral Bone Disorder.Edward R. SmithTim D. HewitsonMichael M. X. CaiParisa AghagolzadehMatthias BachtlerAndreas PaschStephen G. HoltNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-17 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Edward R. Smith Tim D. Hewitson Michael M. X. Cai Parisa Aghagolzadeh Matthias Bachtler Andreas Pasch Stephen G. Holt A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
description	Abstract Calciprotein particles, nanoscale aggregates of insoluble mineral and binding proteins, have emerged as potential mediators of phosphate toxicity in patients with Chronic Kidney Disease. Although existing immunochemical methods for their detection have provided compelling data, these approaches are indirect, lack specificity and are subject to a number of other technical and theoretical shortcomings. Here we have developed a rapid homogeneous fluorescent probe-based flow cytometric method for the detection and quantitation of individual mineral-containing nanoparticles in human and animal serum. This method allows the discrimination of membrane-bound from membrane-free particles and different mineral phases (amorphous vs. crystalline). Critically, the method has been optimised for use on a conventional instrument, without the need for manual hardware adjustments. Using this method, we demonstrate a consistency in findings across studies of Chronic Kidney Disease patients and commonly used uraemic animal models. These studies demonstrate that renal dysfunction is associated with the ripening of calciprotein particles to the crystalline state and reveal bone metabolism and dietary mineral as important modulators of circulating levels. Flow cytometric analysis of calciprotein particles may enhance our understanding of mineral handling in kidney disease and provide a novel indicator of therapeutic efficacy for interventions targeting Chronic Kidney Disease-Mineral Bone Disorder.
format	article
author	Edward R. Smith Tim D. Hewitson Michael M. X. Cai Parisa Aghagolzadeh Matthias Bachtler Andreas Pasch Stephen G. Holt
author_facet	Edward R. Smith Tim D. Hewitson Michael M. X. Cai Parisa Aghagolzadeh Matthias Bachtler Andreas Pasch Stephen G. Holt
author_sort	Edward R. Smith
title	A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
title_short	A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
title_full	A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
title_fullStr	A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
title_full_unstemmed	A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
title_sort	novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/4660db06f2274211a15dd40111002799
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A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum

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