Overview

Explore new frontiers in EV biology

Whether it’s precious clinical samples or hard-to-collect biofluids from developmental biology model organisms, researchers are increasingly challenged by the need to isolate EVs from quite limited sample volumes. To support these researchers, SBI has configured our powerful SmartSEC™ technology into the first commercially available kit optimized for isolating EVs from 10 – 100 µL of biofluid (Table 1). Validated for samples from Drosophila, Planaria, and Arabidopsis, as well as human, mouse, and rat, the SmartSEC™ Mini EV Isolation System is ready to help you push the boundaries of EV biology.

Table 1. SmartSEC Mini recommended input volumes

SmartSEC Mini is a proprietary chromatography-based EV isolation technology that combines all the benefits of size exclusion chromatography (SEC)—purity, yield, reproducibility, and preservation of EV integrity—with a contaminant trapping feature that overcomes the limitations of conventional SEC for fast, easy, and low sample-volume EV isolation.

• The only commercially available EV isolation kit for sample volumes as low as 10 µL
• Great for isolating EVs from a range of biofluids
• Validated in samples from human as well as model developmental biology organisms such as:
  • Drosophila melanogaster
  • Planaria
  • Arabidopsis thaliana
  • Mouse
  • Rat
• Delivers better purity and yield than ultracentrifugation
• Compatible with most downstream applications such as mass spectrometry, western blotting, nanoparticle tracking analysis (NTA), and transmission electron microscopy (TEM)

Each SmartSEC Mini kit comes with pre-loaded SmartSEC Mini columns, collection tubes, and sufficient column buffer for processing 10 samples (see Table 1 for loading volumes).

How It Works

The SmartSEC Mini workflow is quick and easy (Figure 1). Simply pre-wash the SmartSEC Mini column, load your sample, incubate at room temperature for 30-minutes, and centrifuge for 30-seconds at 500 x g to recover isolated EVs. To maximize sample recovery, simply add more buffer to the column and collect up to two additional fractions.

Figure 1. The easy SmartSEC Mini workflow.

Supporting Data

See how well SmartSEC Mini isolates EVs from low volumes in a range of species, biofluids

SmartSEC Mini provides excellent purification of EVs from as little as 10 µL of hemolymph from Drosophila melanogaster, as demonstrated by western blot analysis of 1 µg of equivalent protein (Figure 2). Common Drosophila exosome markers are shown next to molecular weight markers.

Figure 2. SmartSEC Mini provides robust isolation of EVs from Drosophila starting from 10 µL of hemolymph. Data courtesy of Dr. Karen Linnemannstöns, University of Göttingen.

SmartSEC Mini has also been used to isolate EVs from 100 µL Planaria neoblast culture media (Figure 3) and 50 µL of apoplastic fluid from Arabidopsis thaliana (Figure 4) as seen in TEM.

Figure 3. SmartSEC Mini provides robust isolation of EVs from 100 µL of Planaria neoblast culture media. Data courtesy of Dr. Vidyanand Sasidharan, Stowers Institute for Medical Research.

Figure 4. SmartSEC Mini provides robust isolation of EVs from 50 µL of apoplastic fluid from Arabidopsis thaliana. Sample courtesy of Dr. Claudia Uhde-Stone, California State University, East Bay.

SmartSEC Mini provides robust isolation of EVs from 10 µL of mouse serum (Figure 5A, Table 2) and 10 µL of human serum (Figure 5B, Table 2) as demonstrated through western blot data. In addition, SmartSEC Mini delivers more EVs than a competitor’s SEC-based EV isolation product (Table 3).

Figure 5. SmartSEC Mini delivers robust isolation of EVs from human and mouse serum.

SmartSEC Mini specifically enriches for EVs versus lipoprotein particles and blood microparticles as demonstrated via mass spectrometry (Figure 6 and Table 4). We used the quick and easy SmartSEC Mini workflow to isolate EVs from 20 µL of human serum and analyzed the EV-associated proteins via mass spec. Of the ~600 proteins we identified (see Table 4 for a representative list), a functional enrichment analysis conducted using the FunRich tool shows that over 50% of the proteins are known to be associated with exosomes whereas <30% are associated with blood microparticles and <10% are associated with lipoprotein particles (Figure 6).

Figure 6. Mass spectrometry analysis demonstrates that SmartSEC Mini preferentially isolates EVs.

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