evGAG - Performance of evGAG technology in urine
Urine samples were processed with evGAG. Briefly, 1 mL of urine from healthy donor (UHD) and 1 mL of
urine from healthy donor spiked-in (USP) with extracellular vesicles purified from colon cancer cell line
containing KRASG13D mutation (cat nr. EXO-REF-KRAS-G13D-2) were incubated with 2 mL of evGAG each,
for 5 minutes and then centrifuged at 3,000g for 15 minutes. This results in a precipitated pellet containing
Western Blot Analysis of extracellular vesicle markers
Alix, confirmed that the concentration of EVs isolated
by evGAG was higher compared to the competitor.
On the contrary, uromoduline (THP) as not specific
target is less abundant in EVs isolated by evGAG
The pellet containing EVs was resuspended in PBS and analyzed by Nanoparticle Tracking Analysis (NTA).
Nanoparticles concentration is 2 times higher in EVs isolated with evGAG in both UHD and USP samples (8.8
x 1011and 1.2 x 1012respectively) than to EVs isolated with competitor in both UHD and USP samples (4.6 x
1011 and 6.9 x 1011 respectively).
EV RNA was isolated from urine using the extraction phase of SoRTEV™ and analyzed with Bioanalyzer. The
results showed that extracellular vesicles purified with evGAG contain the highest yield of small RNA content
(<200 nt) with a main peak at 100 nt indicating a most efficient extraction of small exoRNA from urine as
oppesed to EVs isolated with competitor kit which showed some longer RNA speciesincluding 18S and 28S
rRNA but a lower peak corresponding to small exoRNA.
Data Sheet (PDF)