Single‐step equipment‐free extracellular vesicle concentration using super absorbent polymer beads

Abstract

Extracellular vesicles (EVs) contain useful biomarkers for disease diagnosis and are promising biomaterials for the delivery of therapeutic molecules in vivo . Accordingly, an efficient concentration method is necessary for large‐scale production or high‐throughput isolation of EVs from bulk liquid samples, including culture medium and body fluids, to achieve their clinical application. However, current EV concentration methods, including ultrafiltration, are limited with respect to cost, efficiency, and centrifugation time. In this study, we developed the first single‐step, equipment‐free EV concentration method using super absorbent polymer (SAP) beads. SAP beads absorb small molecules, including water, via nano‐sized channels but expel and thereby concentrate EVs. Consequently, the beads drastically enrich EVs by reducing the solution volume in a single step, without affecting EV characteristics. Moreover, the purity of the concentrated EV solution was high due to the absorption of protein impurities by SAP beads. To further demonstrate the versatility of the method, we showed that SAP beads successfully enrich EVs in human urine samples and culture medium, enabling better isolation performance than conventional ultrafiltration. We believe the newly developed approach and insight gained in this study will facilitate the use of EVs as prominent biomaterials for disease diagnosis and therapy.

Antioxidative Effects of Carrot-Derived Nanovesicles in Cardiomyoblast and Neuroblastoma Cells

“…Ultracentrifugation provides low yields, as a large number of EVs are lost during centrifugation. In addition, nanovesicles can be disrupted during isolation due to high centrifugal forces, which may complicate the use of isolated nanovesicles as therapeutic drugs [ 23 , 44 , 45 ]. In our previous study, nanovesicles from cabbages were successfully isolated using size-exclusion chromatography combined with ultrafiltration [ 14 ].…”

“…In humans, EVs participate in intercellular communication by transferring biologically active cargoes, including proteins, RNA, DNA, and lipids [14][15][16][17][18]. In this context, EVs produced from mammalian cell cultures have been extensively studied as potential therapeutics or drug delivery vehicles for disease treatment [19][20][21][22][23][24][25]. However, there are crucial limitations to using cell-culture-derived EVs.…”

“…However, there are crucial limitations to using cell-culture-derived EVs. Because a large number of cell-culture-derived EVs are necessary for clinical application, for disease treatment [19][20][21][22][23][24][25]. However, there are crucial limitations to using cell-culturederived EVs.…”

Nanotechnology in emerging liquid biopsy applications

“…The authors succeeded cancer cell enrichment ratios about 5-8 times higher than those provided by untreated surface after a 3-day culture procedure [206]. Absorbent polymer beads (poly(acrylamide-co-acrylic acid)) were also used to provide concentration of exosomes with high purity [207]. Finally, cationic lipid-polymer hybrid nanoparticles were utilized in combination with CHA for the detection of extracellular vesicles (EVs) (LOD = 37.5 particles/mL) [208].…”

Improving Isolation of Extracellular Vesicles by Utilizing Nanomaterials

“…The integrated double-filtration microfluidic device can successfully enrich EVs with a size range of 30-200 nm from urine and has been applied to the study of bladder cancer [92]. Moreover, hydrogels, or forming a hydrogel-like structure, can either exclude EVs or entrap EVs according to the size of their structure and, thereby, achieve EV isolation [93,94]. For instance, super absorbent polymer (SAP) is one hydrogel which has been used as an alternative to filtration for concentrating microorganisms [95].…”