Methods for extracellular vesicle isolation and selective removal
Disclosed is a method for the isolation of extracellular vesicles, including exosomes, from a liquid sample, the method comprising the steps of: adjusting the pH of a liquid sample comprising extracellular vesicles to a preselected, binding pH; contacting the liquid sample with silicon carbide, wherein at the preselected, binding pH, the extracellular vesicles bind to the silicon carbide; and eluting the bound extracellular vesicles from the silicon carbide. The liquid samples can comprise bodily fluids. Further disclosed is a method for producing a liquid sample, substantially depleted of extracellular vesicles, including exosomes.
1. A method for the isolation of extracellular vesicles from a liquid sample, the method comprising the steps of:
(a) adjusting the pH of a liquid sample comprising extracellular vesicles to a preselected, binding pH;
(b) contacting the liquid sample with silicon carbide, wherein at the preselected, binding pH, the extracellular vesicles bind to the silicon carbide; and
(c) eluting the bound extracellular vesicles from the silicon carbide;
wherein steps (a) and (b) can be carried out in any order or simultaneously.
2. The method of claim 1 , wherein the isolated extracellular vesicles comprise exosomes.
3. The method of claim 1 , wherein the liquid sample comprises cell culture media or a biological fluid, wherein the biological fluid is whole blood, blood serum, plasma, urine, saliva, sputum, breast milk, ascetic fluid, semen, vaginal fluid, amniotic fluid, cerebrospinal fluid, sweat or tears.
4. The method of claim 1 , wherein the silicon carbide is in a slurry format or the silicon carbide is packed into a solid support column.
5. The method of claim 4 , wherein the silicon carbide is in a slurry format, and wherein step (b) occurs before step (a), such that the liquid sample is contacted with the silicon carbide before the pH of the liquid sample is adjusted to the preselected binding pH to affect binding of the extracellular vesicles to the silicon carbide.
6. The method of claim 4 , wherein the silicon carbide is in a slurry format, and wherein prior to eluting the bound extracellular vesicles from the silicon carbide, the method further comprises the step of separating the silicon carbide with the bound extracellular vesicles from the liquid sample by centrifugation or by gravity settling.
7. The method of claim 1 , wherein preselected, binding pH is about 7 to about 11.
8. The method of claim 7 , wherein the preselected, binding pH is about 8.5.
9. The method of claim 1 , wherein the bound extracellular vesicles are eluted from the silicon carbide using a low salt buffer having a pH of about 4 to about 6.
10. The method of claim 1 , wherein the isolated extracellular vesicles have an average diameter of about 40 nm to about 150 nm.