Hi,
I have isolated exosomes from plasma, the exosomes are in some sort of protein aggregate. Is there anyway to wash away other plasma proteins (albumins, fibrinogens etc.) without lysing the exosomes? Depletion is not really an option.
Hi Tim, you could try treating the exosomes with trypsin to get rid of the external proteins.
http://www.nature.com/cr/journal/v22/n3/full/cr2011196a.html
http://jvi.asm.org/content/83/2/512.full
I am currently treating with proteinase K but we want to keep surface proteins intact.
Hi Tim, when you say that you want to keep surface proteins intact, do you mean exosome membrane proteins?
If so, you could try TX-114 phase separation of membrane proteins. This treatment can enrich for membrane proteins, but will lyse the exosomes and their contents will be lost
If this is not what you are after, have you tried a discontinuous density sucrose cushion?
.https://clincancerres.aacrjournals.org/content/11/20/7554.full
The purpose is to compare a kit with ultracentrifugation, the key problem are the plasma proteins. The exosomes are captured in a protein aggregate together with, apparantly, lots of plasma proteins. The final purpose is to measure proteins inside the exosomes as well as on the outsidd of the exosomes using lc-ms/ms.
- Badges
- Science method