TY - JOUR
T1 - Sequential Filtration
T2 - A Gentle Method for the Isolation of Functional Extracellular Vesicles
AU - Heinemann, Mitja L.
AU - Vykoukal, Jody
PY - 2017
Y1 - 2017
N2 - A prevalent challenge in isolating extracellular vesicles (EVs) from biological fluids is the reliable depletion of abundant contaminants-including free proteins and biomolecules, as well as nontarget vesicle subpopulations and other nanoparticulates-from the sample matrix while maximizing recovery. Sequential Filtration is a recently published approach for the size-based isolation of exosomes that is ideally suited for large-volume biofluid samples such as ascites , urine , lavage fluid, or cell-conditioned media. We describe a straightforward, three-step protocol comprising back-to-back steps of dead-end (normal) filtration, tangential-flow filtration, and track-etched membrane filtration that can be applied to yield a homogeneous population of exosome-sized extracellular vesicles. The approach is scalable and employs relatively gentle manipulation forces to fractionate and concentrate extracellular vesicles with good purity and functional integrity.
AB - A prevalent challenge in isolating extracellular vesicles (EVs) from biological fluids is the reliable depletion of abundant contaminants-including free proteins and biomolecules, as well as nontarget vesicle subpopulations and other nanoparticulates-from the sample matrix while maximizing recovery. Sequential Filtration is a recently published approach for the size-based isolation of exosomes that is ideally suited for large-volume biofluid samples such as ascites , urine , lavage fluid, or cell-conditioned media. We describe a straightforward, three-step protocol comprising back-to-back steps of dead-end (normal) filtration, tangential-flow filtration, and track-etched membrane filtration that can be applied to yield a homogeneous population of exosome-sized extracellular vesicles. The approach is scalable and employs relatively gentle manipulation forces to fractionate and concentrate extracellular vesicles with good purity and functional integrity.
KW - Cancer biology
KW - Early detection
KW - Exosome isolation
KW - Exosomes
KW - Extracellular vesicles
KW - Functional exosomes
KW - Sequential filtration
UR - http://www.scopus.com/inward/record.url?scp=85041909502&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85041909502&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7253-1_4
DO - 10.1007/978-1-4939-7253-1_4
M3 - Article
C2 - 28828646
AN - SCOPUS:85041909502
SN - 1064-3745
VL - 1660
SP - 33
EP - 41
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -