Extracellular vesicle isolation from urine: Exo-Urine EV Isolation Kit

The innovative Exo-Urine™ EV Isolation Kit from System Biosciences enables the isolation of high-purity extracellular vesicles (EVs) from urine in less than one hour. With minimal carryover of THP and other proteins and high EV yield, the Exo-Urine EV Isolation Kit enables more efficient biomarker discovery from urine-derived EVs.

Benefits of using Exo-Urine EV Isolation Kit
• High purity EVs isolated
• High yield of EVs
• Minimal carryover of THP and other proteins
• Isolate EVs from 10 samples in less than one hour
• Low input sample volume
• Utilises fast-flow resin/gravity column
• Compatible with fresh and frozen urine
• Sensitive isolation method – decreased contaminants and enrichment of EV-associated markers compared with column, precipitation, and ultracentrifugation isolation methods

Urinary EVs (uEVs) were enriched using Exo-Urine EV Isolation Kit and Company N’s kit from freshly collected pooled uringe samples from three healthy donors and stored at two hrs, four hrs and six hrs at +four degrees C. 1.5 ml urine was used for the Exo-Urine EV Isolation Kit compared with 15 ml for Company N’s kit. Western blot analysis for (A) THP and CD63 comparing the two kits and (B) AQP2, TSG101 AND HSP70 for the Exo-Urine™ EV Isolation Kit demonstrates superior performance of the Exo-Urine EV Isolation Kit for delivering low level contaminants and enrichment of EV-specific markers.

Urinary EVs (uEVs) were enriched from pooled urine samples stored at -200 C for three weeks using Exo-Urine EV Isolation Kit, Company N’s kit, ultracentrifugation and polymer precipitation isolation methods. 1.5 ml urine was used for Exo-Urine EV Isolation Kit compared with 15 ml for the other methods. Different exosomal markers indicated above were probed to define the isolated uEVs from different methods (loaded based on equal volume ratio) using immunoblotting. High relative amount of aquaporin-2 (AQP2) vs other EV markers (in UC/polymer precipitation lanes) indicates possible carryover of non-EV associated soluble form of aquaporin-2 (Kodaka et al. 2018).

The Exo-Urine EV Isolation Kit delivers higher yields of EVs (protein equivalent) than other methods

EV preparations isolated by Exo-Urine, Company N, ultracentrifugation and polymer precipitation were analysed for EV yield, measured by protein equivalence using Qubit™ protein assay, and amounts normalised to the EVs isolated by ultracentrifugation. Use of the Exo-Urine™ EV Isolation Kit results in more EVs than all other methods.

Applications
• Biomarker discovery

Material available for download
Exo-Urine EV Isolation Kit manual
Exosome research products and services brochure

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