Automated Enrichment and Coextraction Pipeline for Efficient Analysis of Urinary Extracellular Vesicle Protein and RNA

Extracellular vesicle (EV) cargoes
have great potential as promising
biomarkers for disease diagnosis. However, traditional methods for
EV enrichment and cargo extraction are inefficient, limiting the accessibility
of valuable biological information. Here, we present a bead-based
pipeline designed to efficiently enrich EVs from urine while simultaneously
extracting both EV protein and RNA. Compared to traditional methods
involving ultracentrifugation (UC) for EV isolation, acetone precipitation
for protein extraction, and TRIzol for RNA extraction, the new pipeline
achieved over a 5-fold improvement in EV protein and RNA yield. Additionally,
it reduced the duration from approximately 2 days to just 1 h. When
applied to the noninvasive diagnosis of prostate cancer (PCa), the
pipeline demonstrated excellent diagnostic performance, with a panel
of EV protein and RNA markers achieving an AUC of 0.82. This efficient
and streamlined pipeline provides a robust tool for comprehensive
EV cargo analysis and holds great potential for advancing noninvasive
diagnostics in clinical settings.