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Rapid isolation and quantification of extracellular vesicles from suspension-adapted human embryonic kidney cells using capillary-channeled polymer fiber spin-down tips.

Exosomes, a subset of extracellular vesicles (EVs, 30-200 nm diameter), serve as biomolecular snapshots of their cell of origin and vehicles for intercellular communication, playing roles in biological processes including homeostasis maintenance and immune modulation. The large-scale processing of exosomes for use as therapeutic vectors has been proposed, but these applications are limited by impure, low-yield recoveries from cell culture milieu (CCM). Current isolation methods are also limited by tedious and laborious workflows, especially towards isolation of EVs from CCM for therapeutic applications. Employed is a rapid (<10 minute) EV isolation method on a capillary-channeled polymer (C-CP) fiber spin-down tip format. EVs are isolated from the CCM of suspension-adapted human embryonic kidney cells (HEK293), one of the candidate cell lines for commercial EV production. This batch solid-phase extraction technique allows 1012 EVs to be obtained from only 100 μL aliquots of milieu, processed using a benchtop centrifuge. The tip-isolated EVs were characterized using transmission electron microscopy (TEM), multi-angle light scattering (MALS), absorbance quantification, an enzyme-linked immunosorbent assay (ELISA) to tetraspanin marker proteins, and a protein purity assay. It is believed that the demonstrated approach has immediate relevance in research and analytical laboratories, with opportunities for production-level scale-up projected. This article is protected by copyright. All rights reserved.