Open AccessAsymmetric depth‐filtration: A versatile and scalable method for high‐yield isolation of extracellular vesicles with low contamination
Author(s) -
Chernyshev Vasiliy S.,
ChuprovNetochin Roman N.,
Tsydenzhapova Ekaterina,
Svirshchevskaya Elena V.,
Poltavtseva Rimma A.,
Merdalimova Anastasiia,
Yashchenok Alexey,
Keshelava Amiran,
Sorokin Konstantin,
Keshelava Varlam,
Sukhikh Gennadiy T.,
Gorin Dmitry,
Leonov Sergey,
Skliar Mikhail
Publication year - 2022
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1002/jev2.12256
Subject(s) - filtration (mathematics) , extracellular vesicles , chromatography , extracellular vesicle , size exclusion chromatography , contamination , isolation (microbiology) , nanoparticle tracking analysis , chemistry , yield (engineering) , ultracentrifuge , exosome , microvesicles , nanotechnology , computer science , materials science , bioinformatics , biology , biochemistry , mathematics , microbiology and biotechnology , enzyme , ecology , microrna , statistics , gene , metallurgy
We developed a novel asymmetric depth filtration (DF) approach to isolate extracellular vesicles (EVs) from biological fluids that outperforms ultracentrifugation and size‐exclusion chromatography in purity and yield of isolated EVs. By these metrics, a single‐step DF matches or exceeds the performance of multistep protocols with dedicated purification procedures in the isolation of plasma EVs. We demonstrate the selective transit and capture of biological nanoparticles in asymmetric pores by size and elasticity, low surface binding to the filtration medium, and the ability to cleanse EVs held by the filter before their recovery with the reversed flow all contribute to the achieved purity and yield of preparations. We further demonstrate the method's versatility by applying it to isolate EVs from different biofluids (plasma, urine, and cell culture growth medium). The DF workflow is simple, fast, and inexpensive. Only standard laboratory equipment is required for its implementation, making DF suitable for low‐resource and point‐of‐use locations. The method may be used for EV isolation from small biological samples in diagnostic and treatment guidance applications. It can also be scaled up to harvest therapeutic EVs from large volumes of cell culture medium.