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

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    Author
    Smith, ER; Hewitson, TD; Cai, MMX; Aghagolzadeh, P; Bachtler, M; Pasch, A; Holt, SG
    Date
    2017-07-18
    Source Title
    Scientific Reports
    Publisher
    NATURE PUBLISHING GROUP
    University of Melbourne Author/s
    Hewitson, Timothy; Smith, Edward; Holt, Stephen; Cai, Michael
    Affiliation
    Medicine and Radiology
    Metadata
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    Document Type
    Journal Article
    Citations
    Smith, E. R., Hewitson, T. D., Cai, M. M. X., Aghagolzadeh, P., Bachtler, M., Pasch, A. & Holt, S. G. (2017). A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum. SCIENTIFIC REPORTS, 7 (1), https://doi.org/10.1038/s41598-017-05474-y.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/256576
    DOI
    10.1038/s41598-017-05474-y
    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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