Evidence map›Paper›PMID 42289649›Full record

ArticleClinical proteomics2026

Comparative mass spectrometry analysis of high and low centrifugation extracellular vesicle (EV) pellets from healthy urine following Tamm-Horsfall protein removal.

Dana Bielopolski, Luca Musante, Henrik Molina, Douglas Barrows, Samantha Upson, Lifang Yang, Thomas Carrol, Jonathan N Tobin, Rhonda G Kost, U Erdbrügger

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Article in Clinical proteomics, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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2 · The registry

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3 · Its place in the literature

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4 · The record

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5 · Who and what money

Authors and funding

10 authors.

Dana BielopolskiThe Rockefeller University Center for Clinical and Translational Science, NY, 1230 York Avenue, New York, 10065, USA. dbielopols@rockefeller.edu.
Luca MusanteExtracellular Vesicle Core, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, USA.
Henrik MolinaProteomics Resource Center, The Rockefeller University, New York, NY, USA.
Douglas BarrowsBioinformatics Resource Center, The Rockefeller University, New York, NY, USA.
Samantha UpsonDepartment of Medicine, Division of Nephrology, University of Virginia at Charlottesville, Charlottesville, USA.
Lifang YangDepartment of Biomedical & Translational Sciences, Eastern Virginia Medical School at Old Dominion University, Norfolk, USA.
Thomas CarrolBioinformatics Resource Center, The Rockefeller University, New York, NY, USA.
Jonathan N TobinThe Rockefeller University Center for Clinical and Translational Science, NY, 1230 York Avenue, New York, 10065, USA.
Rhonda G KostThe Rockefeller University Center for Clinical and Translational Science, NY, 1230 York Avenue, New York, 10065, USA.
U ErdbrüggerDepartment of Medicine, Division of Nephrology, University of Virginia at Charlottesville, Charlottesville, USA.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

objectivesUrinary extracellular vesicles (uEVs) are potential biomarkers for renal physiology and disease. Their isolation is challenged by Tamm-Horsfall protein (THP) contamination, particularly in high-speed ultracentrifugation (P100) fractions. Low-speed (P20) fractions contain biologically relevant uEVs, yet their proteomic composition is underexplored. This study evaluates strategies for THP removal to optimize mass spectrometry analysis and characterizes P20 and P100 uEV proteomes and its combination in healthy urine.

methodsUrine from a healthy volunteer was processed via sequential centrifugation at 21,200 g (P20) and 164,244 g (P100). THP removal was performed using low ionic strength buffer (P20) and size-exclusion chromatography (P100). Western blotting, nanoparticle tracking, and mass spectrometry were used to characterize protein content. Functional analysis was performed to assess biological significance.

resultsMass spectrometry identified 1,103 unique proteins in P100, 929 in P20, and 1,676 in the combined pellet, with 90% similarity between fractions. P20 was enriched in proteasome components (PSMC2, PSMD2, PSMD5, PSMD6, PSMD8, PSMD11, PSMD13, PSME1), while P100 had no advantage in terms of expression profile in any of the proteins examined. Functional analysis indicated immune activation in P100 and protein degradation in P20, suggesting complementary biological roles.

conclusionThis study reveals proteasome enrichment in P20 uEVs and supports a combined P20 + P100 approach to improve biomarker discovery. Optimized THP removal strategies enhance mass spectrometry resolution, facilitating uEV analysis for clinical and research applications. INSTITUTIONAL REVIEW BOARD FOR HEALTH SCIENCES RESEARCH (IRB-HSR) AT UVA: HSR220053.

Indexed as

FractionationProteomicsTamm-Horsfall proteinUrinary extracellular vesicles

Identifiers

PMID42289649
PMCPMC13523286

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.