SmartSEC™ Single for EV Isolation

SmartSEC Single performs better than competitor’s q SEC technology

To understand how well SmartSEC Single performs compared to a competitor’s q SEC columns, we isolated EVs from 250 µL of human serum using both SmartSEC Single and q SEC columns and analyzed 1 µg of protein equivalent from each isolation method on a western blot (Figure 1) and using fluorescent nanoparticle tracking analysis (fNTA, Figure 2). EVs isolated using SmartSEC Single show lower levels of carry-over proteins such as albumin and IgGs (Figure 1) and a higher number of particles per mg of protein (Figure 2) than EVs isolated using the q SEC columns, demonstrating the higher purity delivered by SmartSEC Single.

In addition, SmartSEC Single delivered EVs at ~6-fold higher concentration than the q SEC columns (Figure 1), for overall superior performance.

Figure 1. Western blot analysis shows that SmartSEC Single delivers higher yields of cleaner EVs than a competitor’s q SEC columns.

Figure 2. fNTA show that SmartSEC Single delivers higher yields of cleaner EVs than a competitor’s q SEC column.

SmartSEC Single cleanly isolates EVs from CSF

SmartSEC Single can isolate EVs from cerebrospinal fluid (CSF) with no detectable cellular contamination, as shown in the ExoCheck Neuro Array in Figure 3. The isolated EVs possess both neuronal and exosomal markers, suggesting that the EV prep is enriched for vesicles of neuronal origin. Interestingly, EVs isolated from CSF tend to be smaller in size than EVs isolated from serum (Figure 4).

Figure 3. SmartSEC Single isolates neuronal EVs from CSF.

Figure 4. EVs isolated from CSF by SmartSEC Singles are smaller than EVs isolated from serum.

EVs isolated using SmartSEC Single possess typical EV morphology

Transmission electron microscopy (TEM) of EVs isolated from serum using SmartSEC Single possess typical EV morphology—intact vesicles with a double layer of membranes—with little visible background debris (Figure 5).

Figure 5. EVs isolated using SmartSEC Single possess typical EV morphology.

EVs isolated with SmartSEC Single and a competitor’s q SEC columns have very similar proteomic profiles

We analyzed EVs isolated from human serum using SmartSEC Single and a competitor’s q SEC columns using mass spectrometry. Both methods yielded very similar proteomes, with 86% overlap (Figure 6).

Figure 6. EVs isolated with SmartSEC Single and a competitor’s q SEC columns have very similar proteomic profiles