A Rapid Exosome Isolation Using Ultrafiltration and Size Exclusion Chromatography (REIUS) Method for Exosome Isolation from Melanoma Cell Lines

Shu, Shin La; Allen, Cheryl; Benjamin-Davalos, Shawna; Koroleva, Marina; MacFarland, Don; Minderman, Hans; Ernstoff, Marc S.

doi:10.1007/978-1-0716-1205-7_22

Cited by 20 publications

(13 citation statements)

References 15 publications

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“…S3). In summary, these results confirm work by others 14,[31][32][33][34][35][36][37][38][39] that the size exclusion column methods of isolation yield pure EV for MASEV analysis in reasonable amounts of time.…”

Section: Optimization Of Masevsupporting

confidence: 89%

Multiplexed analysis of a single EV (MASEV) reveals unique biomarker composition with diagnostic impact

Spitzberg

Ferguson

Yang

et al. 2022

Preprint

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Exosomes and extracellular vesicles (EV) are increasingly being explored as circulating biomarkers, but their heterogenous composition will likely mandate the development of single EV technologies. Highly multiplexed analyses of single EVs have been challenging to implement beyond a few colors during spectral sensing. We use a multiplexed analysis of the single EV technique (MASEV) to interrogate thousands of individual EVs during 5 cycles of multi-channel fluorescence staining for 15 EV biomarkers. Contrary to the common belief, we show that i) several “ubiquitous” markers are much less common than believed; ii) that multiple biomarkers concur in single vesicles but only in small fractions, iii) that affinity purification can lead to loss of rare EV subtypes, and iv) that deep profiling allows detailed analysis of EV potentially improving the diagnostic content. These findings establish the potential of MASEV for uncovering fundamental EV biology and heterogeneity and increasing diagnostic specificity. Abstract Figure Graphical abstract Multiplexed analysis of single EV (MASEV) allows robust protein profiling at the single EV level, a prerequisite for early cancer detection or organ of origin determination.

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Section: Optimization Of Masevsupporting

confidence: 89%

Multiplexed analysis of a single EV (MASEV) reveals unique biomarker composition with diagnostic impact

Spitzberg

Ferguson

Yang

et al. 2022

Preprint

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“…To improve the isolation purity, researchers have tried to combine ultracentrifugation and ultrafiltration with size-exclusion chromatography for the isolation of exosomes (Guan et al, 2020;Shu et al, 2021;Turner et al, 2022). Studies have shown that ultrafiltration combined with size-exclusion chromatography can reduce the level of impurity cytokine (interleukin-10) in isolated exosomes.…”

Section: Improvements Of Traditional Methods For Exosome Isolationmentioning

confidence: 99%

Recent developments in isolating methods for exosomes

Gao

et al. 2023

Front. Bioeng. Biotechnol.

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Exosomes are the smallest extracellular vesicles that can be released by practically all cell types, and range in size from 30 nm to 150 nm. As the major marker of liquid biopsies, exosomes have great potential for disease diagnosis, therapy, and prognosis. However, their inherent heterogeneity, the complexity of biological fluids, and the presence of nanoscale contaminants make the isolation of exosomes a great challenge. Traditional isolation methods of exosomes are cumbersome and challenging with complex and time-consuming operations. In recent years, the emergence of microfluidic chips, nanolithography, electro-deposition, and other technologies has promoted the combination and innovation of the isolation methods. The application of these methods has brought very considerable benefits to the isolation of exosomes such as ultra-fast, portable integration, and low loss. There are significant functional improvements in isolation yield, isolation purity, and clinical applications. In this review, a series of methods for the isolation of exosomes are summarized, with emphasis on the emerging methods, and in-depth comparison and analysis of each method are provided, including their principles, merits, and demerits.

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“…Shu et al rapidly separated exosomes derived from human melanoma (HMEX) by ultrafiltration and then demonstrated enrichment using molecular and microscopic methods. 72 This new method for extracting cellular exosomes is more effective with low-speed centrifugation and could reduce the damage caused by pressure on exosomes. 73…”

Section: Ultrafiltration Methodsmentioning

confidence: 99%