Chapter title |
Sequential Filtration: A Gentle Method for the Isolation of Functional Extracellular Vesicles
|
---|---|
Chapter number | 4 |
Book title |
Extracellular Vesicles
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7253-1_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7251-7, 978-1-4939-7253-1
|
Authors |
Mitja L. Heinemann, Jody Vykoukal, Heinemann, Mitja L., Vykoukal, Jody |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 63 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 13 | 21% |
Student > Ph. D. Student | 7 | 11% |
Student > Master | 6 | 10% |
Student > Bachelor | 4 | 6% |
Student > Doctoral Student | 3 | 5% |
Other | 8 | 13% |
Unknown | 22 | 35% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 10 | 16% |
Medicine and Dentistry | 9 | 14% |
Agricultural and Biological Sciences | 5 | 8% |
Engineering | 5 | 8% |
Chemical Engineering | 3 | 5% |
Other | 5 | 8% |
Unknown | 26 | 41% |