Workflow and Performance
How does EXO-NET® work?
Workflow for EV capture
EXO-NET® bead volumes may be adjusted according to biofluid or application. However, the following protocol may be applied for most generic applications enabling EVs isolation from body fluids within 15 minutes for downstream analysis of EV biomarkers (proteins, RNAs, lipids).
This workflow can be adapted for high throughput screening of EVs and exosome-based biomarker discovery for diagnostics and therapeutic targets.
Performance: EV Capture
In comparison testing, EXO-NET® successfully enriched plasma EVs and outperformed competitor products.
EXO-NET® - High Yield and Capture of EVs from Various Biofluids
ZetaView analysis of Normal Human Plasma (NHP) depleted by EV capture kits. 500 uL NHP was incubated with 30 uL EXONET for 15 minutes before separating the EXONET bounds EVs by using the magnetic rack. Similarly, EVs from 500 uL NHP plasma were isolated by two comparable bead-based kits. The input NHP along with the plasma depleted with EXONET, bead-based kits were analyzed using a ZetaView nanoparticle tracking analyser. (A) ZetaView profile of particle counts in input and depleted plasma against particle size is plated. (B) % particles captured by different EV capture tools are plotted. Bars indicate SEM (n= 3). The EV captured by comparable bead-based kits are highlighted (bottom panel).
Performance: Purity and enrichment of plasma EV protein markers
Higher Summed Peptide Intensity of Albumin in Competitor Beads
Higher Summed Peptide Intensity of Exosomal Protein Markers in EXO-NET®
EV Protein Markers
EXO-NET® Pab-Exosome Capture results in enrichment of EV Proteins compared to Size Exclusion Chromatography
EV Protein Markers
EXO-NET® Pab-Exosome Capture demonstrates higher enrichment (Panel A) and reduced co-isolated of contaminates (Panel B) compared to other isolation kits tested.
Performance: efficient isolation of plasma EV RNA markers
EV lysates isolated using EXO-NET® results in equivalent or higher recovery of EV RNA compared to 4 commercial EV isolation kits as indicated by lower CT values. mRNA assayed by RT-qPCR for GAPDH and OAZ1, RPLP0 and SERF2.
EV lysates isolated using EXO-NET and 4 other competitor products were assayed by RT-qPCR for miRNAs miR-16, let-7a and miR-21.