evGAG, extracellular visicles purification kit

evGAG is a patented purification method that allows isolation of extracellular vesicles (EVs) from biofluids.

The evGAG reaction is based on the interaction between the precipation solution and glycosaminoglycans

(GAGs) in the EVs. The product is ideal for the discovery of EV associated biomarkers.

Applications

• Ideal for Isolation of extracellular vesicles (EVs) from urine samples

• Nanoparticle Tracking Analysis (NTA)

• Flow Cytometry Analysis (FACS)

• Western Blot (WB)

Advantages

• High sensitivity and specificity

• Rapid and simple process (20 minutes)

• Inexpensive

• Small sample needed

• No specific equipment needed

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

EVs.

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)