Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties

Nordin, Joel Z.; Lee, Yi; Vader, Pieter; Mäger, Imre; Johansson, Henrik J.; Heusermann, Wolf; Wiklander, Oscar P. B.; Hällbrink, Mattias; Seow, Yiqi; Bultema, Jarred J.; Gilthorpe, Jonathan; Davies, T.G.; Fairchild, Paul J.; Gabrielsson, Susanne; Meisner‐Kober, Nicole; Lehtiö, Janne; Smith, C. I. Edvard; Wood, Matthew; Andaloussi, Samir El

doi:10.1016/j.nano.2015.01.003

Cited by 501 publications

(460 citation statements)

References 18 publications

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“…As EVs are abundantly present and stable in the blood circulation, it was speculated that EVs could have longer circulation times and mediate drug targeting to extrahepatic and non-lymphoid tissues. However, reports studying the PK of IV injected EVs described short half-lives (~2 minutes [115,116] and ~20 minutes [117]) with predominant uptake by liver, lung, kidney and spleen, thus closely resembling the biodistribution of synthetic liposomes [70,118,119]. The elimination after IV injection occurs via hepatic and renal routes [117] in which MPS-associated macrophages seem to play a key role [118].…”

Section: Extracellular Behavior Of Evsmentioning

confidence: 98%

“…Additionally, the impact of the high shear forces on the vesicle integrity are under debate. While some studies indicate no changes in the integrity of the EVs after UC [68,69] others show subtle EV aggregation influencing the EV biodistribution [70,71].…”

Section: Ev Purification Protocols and Stabilitymentioning

confidence: 99%

“…SEC enables fast sample processing with high yield making it applicable in a clinical setting [70,84]. However, sample dilution is inherent to the technique as well as co-purification of (a low percentage of) lipoproteins, which might limit the applicability of this technique in a diagnostic context [74,83].…”

Section: Ev Purification Protocols and Stabilitymentioning

confidence: 99%

“…Furthermore, also the selected purification protocol or the transfer of allogeneic EVs can potentially influence the EV's PK profile [70].…”

Section: Extracellular Behavior Of Evsmentioning

confidence: 99%

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Therapeutic and diagnostic applications of extracellular vesicles

Stremersch

Smedt

Raemdonck

2016

Journal of Controlled Release

152

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During the past two decades, extracellular vesicles (EVs) have been identified as important mediators of intercellular communication, enabling the functional transfer of bioactive molecules from one cell to another. Consequently, it is becoming increasingly clear that these vesicles are involved in many (patho)physiological processes, providing opportunities for therapeutic applications. Moreover, it is known that the molecular composition of EVs reflects the physiological status of the producing cell and tissue, rationalizing their exploitation as biomarkers in various diseases. In this review the composition, biogenesis and diversity of EVs is discussed in a therapeutic and diagnostic context. We describe emerging therapeutic applications, including the use of EVs as drug delivery vehicles and as cell-free vaccines, and reflect on future challenges for clinical translation. Finally, we discuss the use of EVs as a biomarker source and highlight recent studies and clinical successes.

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Section: Extracellular Behavior Of Evsmentioning

confidence: 98%

Section: Ev Purification Protocols and Stabilitymentioning

confidence: 99%

Section: Ev Purification Protocols and Stabilitymentioning

confidence: 99%

“…Furthermore, also the selected purification protocol or the transfer of allogeneic EVs can potentially influence the EV's PK profile [70].…”

Section: Extracellular Behavior Of Evsmentioning

confidence: 99%

See 2 more Smart Citations

Therapeutic and diagnostic applications of extracellular vesicles

Stremersch

Smedt

Raemdonck

2016

Journal of Controlled Release

152

View full text Add to dashboard Cite

show abstract

“…However, the promotion of this method which required the ultracentrifugation step is limited in ordinary labs due to the high-cost centrifuge equipment. More importantly, the tough process during ultracentrifugation may also induce exosomes to aggregate [14] or crack [15]. To overcome the above shortcomings, we developed a facile and low cost method for highly-purified urinary exosomes isolation by using 300 kDa dialysis tubes and without ultracentrifugation.…”

Section: Introductionmentioning

confidence: 99%

High-purified Isolation and Proteomic Analysis of Urinary Exosomes from Healthy Persons

Yang¹,

Zhi²,

Liu³

et al. 2017

Nano BioMed ENG

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Urinary exosomes containing specific biomarkers have recently been considered as novel potential non-invasive candidates for renal disease diagnosis. However, the development of urinary exosomes in basic research and their subsequent diagnostic application are impeded by the lack of an efficient isolation method. One of the main challenges during urinary exosomes isolation is how to remove a large number of Tamm Horsfall proteins (around 92 kDa) and other biological components from exosome enrichment mixture. Herein, we report a facile and low-cost isolation method for highlypurified human urinary exosomes based on dialysis. The key protocol for exosome isolation includes only two steps: (1) Healthy person urines were collected. 10 mL urine in 300 kDa dialysis tubes was firstly dialyzed in phosphate-buffered saline solution three times for sequential nine hours; (2) The dialysis suspension was concentrated to 200 μL by using 100 kDa ultracentrifuge tubes to achieve urinary exosome isolation. For verification, the concentrated solution was examined by western blot, transmission electronic microscopy, atomic force microscopy and qNano, which demonstrated the highly-purified urinary exosomes were present. Furthermore, a total of 359 proteins were identified by the proteomic analysis of purified urinary exosomes from healthy persons. Those results demonstrated that highly-purified urinary exosomes could be achieved by our isolation method; 359 proteins were identified from healthy persons. Further works will focus on screening and identifying disease-related biomarkers from human urine exosomes for clinical diagnosis.

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Exosome‐associated polysialic acid modulates membrane potentials, membrane thermotropic properties, and raft‐dependent interactions between vesicles

et al. 2020

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In mammals, polysialic acid (polySia) attached to a small number of transmembrane protein carriers occurs on the surface of plasma membranes of neural, cancer, immune, and placental trophoblast cells. Here, our goal was to demonstrate the presence of polySia on exosomes and its effect on membrane properties. We isolated exosomes and found that polysialylated exosomes in fetal bovine serum originate mostly from placental trophoblasts, while in calf bovine serum, they originate from immune cells. Enzymatic removal of polySia chains from the exosomal surface makes the membrane surface potential more positive, transmembrane potential more negative, and reduces the activation energy for membrane anisotropy changes. We demonstrate for the first time that exosomes could interact through polySia-raft interactions. We suggest that polysialylation of exosomal membrane can have a thermo-protecting effect and can modulate exosome-plasma membrane interactions.Exosomes are small extracellular vesicles (EVs), 50 Ä 150 nm in diameter, that originate from the endosome and are released from cells when multivesicular bodies (MVBs) containing intraluminal vesicles (ILVs) fuse with the plasma membrane. Microvesicles (100 Ä 1000 nm in diameter) are larger than exosomes and are released from cells through blebbing (budding out) and fission of the plasma membrane. There is a small overlap of exosomes and microvesicles concerning their size; however, vesicles within the diameter range of 50 Ä 100 nm are considered as exosomes. In addition to exosomes and microvesicles, there are also other nonvesicular nanoparticles in the body fluids, for example, lipoproteins and their complexes [1]. Exosomes are mainly recovered after high-speed ultracentrifugation, but such preparations may include the smallest non-EV structures, for example, high-density lipoprotein (5 Ä 35 nm in diameter) [1]. However, lipoproteins can be separated from exosomes by gel filtration method using appropriate matrix [2,3]. Exosomes are involved in mammalian pathophysiology including cancer, immune responses, cardiovascular diseases, regeneration, and stem cell-based therapy (reviewed in ref. [4]). Exosomes in cancer seem to be Abbreviations ANS, 8-anilino-1-naphthalenesulfonic acid ammonium salt

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Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties

Cited by 501 publications

References 18 publications

Therapeutic and diagnostic applications of extracellular vesicles

Therapeutic and diagnostic applications of extracellular vesicles

High-purified Isolation and Proteomic Analysis of Urinary Exosomes from Healthy Persons

Exosome‐associated polysialic acid modulates membrane potentials, membrane thermotropic properties, and raft‐dependent interactions between vesicles

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