qEV EXOSOME ISOLATION

EXOSOME ISOLATION 2

Isolate Exosomes in just 15 minutes.

Izon qEV SEC (size exclusion chromatography) columns provide the most rapid and effective exosome/EV isolation system available. Using a qEV takes around 15 minutes to get a pure sample of intact vesicles, significantly purer than is possible with concentration methods as ultra-centrifugation (UC) or precipitation reagent kits. The resulting samples containing extracellular vesicles (EVs) are consistent, standardisable and repeatable, which is required for both research and clinical testing. qEV SEC columns are the new Gold Standard for exosome/EV isolation. 

 

qEV based separation of exosomes and other EVs offers researchers and clinicians with the following benefits:  

 

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Exosome Isolation 3Prior to the advent of SEC columns, UC was regarded as the gold standard for EV concentration and isolation. UC is time consuming and cannot provide a consistent standardised sample. The very high forces in UC, up to 200,000g, affect and disrupt the vesicles, which may in turn invalidate much of the research. Moreover, aggregated proteins and nucleic acid contaminants are present in the pellet containing the EVs. Density gradient centrifugation (DGC) aims to improve the purity of UC derived exosomes but increases the complexity and time of the procedure even further. Standardised sampling across different laboratories is not achievable with either UC or DGC.

Exosome Isolation Data 

Precipitation reagent kits were adapted for exosome/EV use fromvirus preparation. They are typically PEG based and supposedly sediment the exosomes only.  Published independent data indicates these fractions are heavily contaminated with non-EV material because these reagents cause co-precipitation of proteins, lipoproteins and other biological components. The leading researchers in the EV field no longer use or recommend these products. qEV SEC columns provide clean samples without affecting the structure or function of the EV. Reagent kits provide dirty samples that vary from batch to batch and are no longer recommended for exosome purification.

Izon qEVs isolate the exosomes and other EVs from plasma, conditioned culture media, and other biofluids using SEC. qEV columns come in two different sizes. The qEVoriginal  has a recommended sample size of 0.5 mL and can be cleaned and reused. The qEVsingle is targeted at clinical samples with recommended volume of 100-125mL and is single use only. There are 2 different holders that can simplify and speed up the work even further.

 

 Column Specifications:

Izon Column
qEVoriginal
qEVsingle
Buffer
PBS
PBS
Bed volume (ml)
10
3.5
Sample loading (µl)
250-500
100-150
Fraction volume (µl)
500
200
Exosome elution
Fractions 7-9
Fractions 6-8

 

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References

  1. Gámez-Valero, A., Monguió-Tortajada, M., & Carreras-Planella, L. (2016). Size-Exclusion Chromatography-based isolation minimally alters Extracellular Vesicles’ characteristics compared to precipitating agents. Scientific Reports, 6.
  2. Lobb, R. J., Becker, M., Wen Wen, S., Wong, C. S., Wiegmans, A. P., Leimgruber, A., & Möller, A. (2015). Optimized exosome isolation protocol for cell culture supernatant and human plasma. Journal of extracellular vesicles4(1), 27031.
  3. Vanni, I., Alama, A., Grossi, F., Dal Bello, M. G., & Coco, S. (2017). Exosomes: a new horizon in lung cancer. Drug Discovery Today.
  4. Mateescu, B., Kowal, E. J., van Balkom, B. W., Bartel, S., Bhattacharyya, S. N., Buzás, E. I., ... & Driedonks, T. A. (2017). Obstacles and opportunities in the functional analysis of extracellular vesicle RNA–an ISEV position paper. Journal of Extracellular Vesicles, 6(1), 1286095.
  5. Böing, A. N., Van Der Pol, E., Grootemaat, A. E., Coumans, F. A., Sturk, A., & Nieuwland, R. (2014). Single-step isolation of extracellular vesicles by size-exclusion chromatography. Journal of extracellular vesicles, 3.
  6. van Eijndhoven, M. A., Zijlstra, J. M., Groenewegen, N. J., Drees, E. E., van Niele, S., Baglio, S. R., ... & de Menezes, R. X. (2016). Plasma vesicle miRNAs for therapy response monitoring in Hodgkin lymphoma patients. JCI insight, 1(19).