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oai:repisalud.isciii.es:20.500.12105/272282026-02-13T01:20:18Zcom_20.500.12105_19604com_20.500.12105_2051col_20.500.12105_19605

00925njm 22002777a 4500 dc Zubiri, Irene author Posada-Ayala, Maria author Sanz-Maroto, Aroa author Calvo, Enrique author Martin-Lorenzo, Marta author Gonzalez-Calero, Laura author de la Cuesta, Fernando author Lopez, Juan A author Fernandez-Fernandez, Beatriz author Ortiz, Alberto author Vivanco, Fernando author Alvarez-Llamas, Gloria author 2014-01-16 Diabetic nephropathy (DN) is a major complication of diabetes mellitus (DM), the most frequent cause of end-stage renal disease (ESRD). Exosomes isolated from urine are considered a rich non-invasive source of markers for renal events. Proteinuria associated with DN patients at advanced stages may result in "contamination" of exosomal fraction by co-precipitation of high abundance urine proteins, making it enormously difficult to obtain a reliable comparison of healthy individuals and DN patients and to detect minor proteins. We evaluated different protocols for urinary exosome isolation (ultracentrifugation-based and Exoquick® reagent-based) in combination with an easy and quick depletion procedure of contaminating high abundance proteins (albumin). The optimal methodology was then applied to investigate the proteome of human urinary exosomes in DN and controls using spectral counting LC-MS/MS analysis followed by selected reaction monitoring (SRM) confirmation. A panel of 3 proteins (AMBP, MLL3, and VDAC1) is differentially present in urinary exosomes from DN patients, opening a new field of research focused on improving diagnosis and follow-up of this pathology. J Proteomics. 2014 Jan 16:96:92-102. Journal of Proteomics 24211404 https://hdl.handle.net/20.500.12105/27228 Depletion Diabetic nephropathy Exosomes Label-free quantitation Proteomics Selected reaction monitoring Diabetic nephropathy induces changes in the proteome of human urinary exosomes as revealed by label-free comparative analysis.