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Urinary proteomics sample preparation of proteinuria in exosomes by albumin depletion using AlbuSorb™ Nov. 18, 2013 – MONMOUTH JUNCTION, N.J. — Urine exosome isolation via ultracentrifugation followed by albumin depletion provides researchers with knowledge of renal regulation and could lead to the identification of biomarkers for diabetic nephropathy and diabetic mellitus. By removing albumin using AlbuSorb™ authors identified more urinary proteins such as flotillin-2, lamp-1, PODXL, tsg-101 from exosome fractions of urine samples. Authors Zubiri et al published an article in the Journal of Proteomics which cites AlbuSorb™ for high abundance protein depletion. Titled, ‘Diabetic nephropathy induces changes in the proteome of human urinary exosomes as revealed by label-free comparative analysis’, the article cites protocols of proteomic workflows for biomarker discovery involving isolating exosomes from urine. Proteinuria in urine samples causes contamination of urine samples for proteomics research. Sample preparation protocols with depletion could complement precipitation techniques as less contamination is present in exosomal fractions upon depletion, enrichment, concentration and sample clarification. Urine exosome isolation via ultracentrifugation upon performing depletion provides researchers with knowledge of renal regulation and could lead to the identification of biomarkers for diabetic nephropathy and diabetic mellitus. Upon depletion of high abundance proteins such as albumin, urine exosomes from diabetic and health controls were analyzed by LC-MS/MS and selected reaction monitoring (SRM). The research cites AMBP, MLL3, VDAC1 as proteins in urinary exosomes of diabetic nephropathy patients. AlbuSorb™ is an albumin depletion reagent supplied as a kit with necessary buffers. AlbuSorb™ binds to albumin, and serum proteins flow through. Mild conditions maintain tertiary structure of proteins and simple transfer to secondary analysis. The flow through fraction is compatible with LC-MS, activity based protein profiling and proteomic studies.
Journal of Proteomics detailed protocol Characteristics of AlbuSorb™
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References: Zubiri, Irene, et al. Diabetic nephropathy induces changes in the proteome of human urinary exosomes as revealed by label-free comparative analysis. Journal of Proteomics (2013). *For research use only |
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