Rapid and Efficient Isolation of Exosomes by Clustering and Scattering

Kim, Jinhyun; Lee, Hoyoon; Park, Kyong Hwa; Shin, Sehyun

doi:10.3390/jcm9030650

Cited by 22 publications

(19 citation statements)

References 38 publications

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“…Depending on the incubation time and ambient temperature, the bead volume increased significantly. Similar to the results obtained in a previous study, incubation at 4 °C accelerated the binding of exosomes to the PLL polymer [ 24 ]. Additionally, at 30 min of incubation, the volume increase was fully saturated.…”

Section: Resultssupporting

confidence: 90%

“…Following exosome capture of the bead surface, the zeta potential of the exosome-bound beads became weakly positive (5.4 mV). It is worth noting that the zeta potential of PLL is 42.4 mV, as reported in a previous study [ 24 ]. The strong cationic characteristic of PLL slightly decreased after it bound to the bead and further decreased as the EVs bound to it.…”

Section: Resultssupporting

confidence: 69%

“…The current gold standard isolation method is ultra-centrifugation (UC), which is highly labor-intensive and time-consuming and has a poor yield. Many techniques have been developed for more efficient exosome separation, including size-, charge-, affinity-, and polymer precipitation-based techniques, as well as microfluidics techniques [ 19 , 20 , 21 , 22 , 23 , 24 ].…”

Section: Introductionmentioning

confidence: 99%

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ExoCAS-2: Rapid and Pure Isolation of Exosomes by Anionic Exchange Using Magnetic Beads

Kim

Shin

2021

Biomedicines

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Extracellular vesicles (EVs) are considered essential biomarkers in liquid biopsies. Despite intensive efforts aimed at employing EVs in a clinical setting, workable approaches are currently limited owing to the fact that EV-isolation technologies are still in a nascent stage. This study introduces a magnetic bead-based ion exchange platform for isolating EVs called ExoCAS-2 (exosome clustering and scattering). Owing to their negative charge, exosomes can easily adhere to magnetic beads coated with a polycationic polymer. Owing to the features of magnetic beads, exosomes can be easily processed via washing and elution steps and isolated with high purity and yield within 40 min. The present results confirmed the isolation of exosomes through analyses of size distribution, morphology, surface and internal protein markers, and exosomal RNA. Compared with the commercially available methods, the proposed method showed superior performance in terms of key aspects, including operation time, purity, and recovery rate. This highlights the potential of this magnetic bead-based ion exchange platform for isolating exosomes present in blood plasma.

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Section: Resultssupporting

confidence: 90%

Section: Resultssupporting

confidence: 69%

Section: Introductionmentioning

confidence: 99%

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ExoCAS-2: Rapid and Pure Isolation of Exosomes by Anionic Exchange Using Magnetic Beads

Kim

Shin

2021

Biomedicines

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“…Our data is also relevant for other applications involving EVs, such as isolation and recovery, in vitro EV-cell interaction, etc. 52, 53 and are in agreement with data from virus-containing solutions, where high molecular weight poly-L-lysine (up to 300 KDa) caused relatively higher aggregation than the lower molecular mass polymer polybrene (4-6 kDa) 28 .…”

Section: Discussionsupporting

confidence: 88%

A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma

Campos-Silva

Cáceres‐Martell

Sánchez‐Herrero

et al. 2021

Preprint

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Extracellular vesicles (EVs), released by most cell types, provide an excellent source of biomarkers in biological fluids. Here we describe a method that, using just a few microliters of patient plasma, identifies tumour markers exposed on EVs. Studying physico-chemical properties of EVs in solution, we demonstrate that they behave as stable colloidal suspensions and therefore, in immunocapture assays, many of them are unable to interact with a stationary functionalised surface. Using flocculation methods, like those used to destabilize colloids, we demonstrate that cationic polymers increase EV ζ-potential, diameter, and sedimentation coefficient and thus, allow a more efficient capture on antibody-coated surfaces by both ELISA and bead-assisted flow cytometry. These findings led to optimization of a protocol in microtiter plates allowing effective immunocapture of EVs, directly in plasma without previous ultracentrifugation or other EV enrichment. The method, easily adaptable to any laboratory, has been validated using plasma from lung cancer patients in which the epithelial cell marker EpCAM has been detected on EVs. This high throughput, easy to automate, technology allows screening of large numbers of patients to phenotype tumour markers in circulating EVs, breaking barriers for the validation of proposed EV biomarkers and the discovery of new ones.

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“…Exosomes are different in origin, size, and content and involved in the pathogenesis of infection [ 9 ]. Studies performed over the past years proved that exosomes exist in almost all biological fluids [ 10 , 11 ], including plasma [ 12 , 13 ], serum [ 14 ], bovine and human milk [ 15 ], urine [ 16 ], and saliva [ 17 ]. Importantly, an increasing evidence suggests that exosomes play critical role in cell-to-cell communication [ 18 , 19 ], including the interaction between microorganisms and infected human cells.…”

Section: Introductionmentioning

confidence: 99%

Composition and Clinical Significance of Exosomes in Tuberculosis: A Systematic Literature Review

Biadglegne

König

Rodloff

et al. 2021

JCM

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Tuberculosis (TB) remains a major health issue worldwide. In order to contain TB infections, improved vaccines as well as accurate and reliable diagnostic tools are desirable. Exosomes are employed for the diagnosis of various diseases. At present, research on exosomes in TB is still at the preliminary stage. Recent studies have described isolation and characterization of Mycobacterium tuberculosis (Mtb) derived exosomes in vivo and in vitro. Mtb-derived exosomes (Mtbexo) may be critical for TB pathogenesis by delivering mycobacterial-derived components to the recipient cells. Proteomic and transcriptomic analysis of Mtbexo have revealed a variety of proteins and miRNA, which are utilized by the TB bacteria for pathogenesis. Exosomes have been isolated in body fluids, are amenable for fast detection, and could contribute as diagnostic or prognostic biomarker to disease control. Extraction of exosomes from biological fluids is essential for the exosome research and requires careful standardization for TB. In this review, we summarized the different studies on Mtbexo molecules, including protein and miRNA and the methods used to detect exosomes in biological fluids and cell culture supernatants. Thus, the detection of Mtbexo molecules in biological fluids may have a potential to expedite the diagnosis of TB infection. Moreover, the analysis of Mtbexo may generate new aspects in vaccine development.

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Rapid and Efficient Isolation of Exosomes by Clustering and Scattering

Cited by 22 publications

References 38 publications

ExoCAS-2: Rapid and Pure Isolation of Exosomes by Anionic Exchange Using Magnetic Beads

ExoCAS-2: Rapid and Pure Isolation of Exosomes by Anionic Exchange Using Magnetic Beads

A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma

Composition and Clinical Significance of Exosomes in Tuberculosis: A Systematic Literature Review

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