A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum
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...
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| Published in | Scientific reports Vol. 7; no. 1; pp. 5686 - 17 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
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London
Nature Publishing Group UK
18.07.2017
Nature Publishing Group Nature Portfolio |
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| Online Access | Get full text |
| ISSN | 2045-2322 2045-2322 |
| DOI | 10.1038/s41598-017-05474-y |
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| 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. |
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| AbstractList | 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. 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.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. 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. |
| ArticleNumber | 5686 |
| Author | Holt, Stephen G. Cai, Michael M. X. Pasch, Andreas Smith, Edward R. Hewitson, Tim D. Aghagolzadeh, Parisa Bachtler, Matthias |
| Author_xml | – sequence: 1 givenname: Edward R. orcidid: 0000-0002-2651-1787 surname: Smith fullname: Smith, Edward R. email: edward.smith@mh.org.au organization: Department of Nephrology, The Royal Melbourne Hospital, Department of Medicine - Royal Melbourne Hospital, University of Melbourne – sequence: 2 givenname: Tim D. surname: Hewitson fullname: Hewitson, Tim D. organization: Department of Nephrology, The Royal Melbourne Hospital, Department of Medicine - Royal Melbourne Hospital, University of Melbourne – sequence: 3 givenname: Michael M. X. surname: Cai fullname: Cai, Michael M. X. organization: Department of Nephrology, The Royal Melbourne Hospital, Department of Medicine - Royal Melbourne Hospital, University of Melbourne – sequence: 4 givenname: Parisa surname: Aghagolzadeh fullname: Aghagolzadeh, Parisa organization: Department of Clinical Research, University of Bern – sequence: 5 givenname: Matthias surname: Bachtler fullname: Bachtler, Matthias organization: Department of Clinical Research, University of Bern – sequence: 6 givenname: Andreas surname: Pasch fullname: Pasch, Andreas organization: Department of Clinical Research, University of Bern – sequence: 7 givenname: Stephen G. surname: Holt fullname: Holt, Stephen G. organization: Department of Nephrology, The Royal Melbourne Hospital, Department of Medicine - Royal Melbourne Hospital, University of Melbourne |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28720774$$D View this record in MEDLINE/PubMed |
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| Snippet | Calciprotein particles, nanoscale aggregates of insoluble mineral and binding proteins, have emerged as potential mediators of phosphate toxicity in patients... Abstract Calciprotein particles, nanoscale aggregates of insoluble mineral and binding proteins, have emerged as potential mediators of phosphate toxicity in... |
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| Title | A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum |
| URI | https://link.springer.com/article/10.1038/s41598-017-05474-y https://www.ncbi.nlm.nih.gov/pubmed/28720774 https://www.proquest.com/docview/1956174965 https://www.proquest.com/docview/1920392082 https://pubmed.ncbi.nlm.nih.gov/PMC5515983 https://doaj.org/article/4660db06f2274211a15dd40111002799 |
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