Dear all,
There are a few companies that sell reference EVs, such as the GFP-positive Exosome Standard from Sigma-Aldrich (SAE0193). We have tried to use them in our lab, e.g. to spike them into serum or plasma samples as an internal control. However, the detection of the fluorescent EVs seems to be not very sensitive and a high EV number is required to detect them after EV purification (we use a plate reader).
I was wondering, whether anybody has experience which such EV standards.
Here are my questions:
- What kind of reference EVs have you used and what is your experience?
- How do you detect them?
- For what kind of experiments do you use them?
Thank you very much for your help!
Kathrin
Hi Kathrin,
we have not tried commercially available reference materials so far, however we prepared our own and used them in various methods including single EV flow cytometry (published in Article Optimisation of imaging flow cytometry for the analysis of s...
) and bead-based flow cytometry (Article Systematic Methodological Evaluation of a Multiplex Bead-Bas...
).More recently we also used fluorescently tagged EVs as reference material to evaluate EV stability and storage conditions in various methods including simple bulk plate readers (Article Identification of storage conditions stabilizing extracellul...
)You don't need huge amounts for any of those methods, if you still need some of those EVs I'm happy to share those - just email me :)
Otherwise fully agree there is a high need for more rigorously characterized EV reference materials for different purposes - I know we and some other labs are working on that :)
Hope that helps,
best wishes,
Andre
Hi Kathrin,
We've had good experiences with the HansaBio GFP-CD63 EVs. They label well with anti-CD63 antibody. Concentrations remain consistent.
The Sigma SAE0193 are one to avoid currently. Their current purification process leaves a lot of non-EV material in there. But I expect they will change this in the future as the group they received the patent from were able to produce very good fluorescent EVs.
Our use is for nFCM, we currently use them as spike-ins as proof of ability to label. We also intend to include GFP FL values and APC labelled values as a rudimentary reference. Lot variation is going to limit their usefulness in this regard but options are still limited currently. I'd love to hear of other reference materials.
Cheers,
Ben
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