High-throughput exosome isolation based on bidirectional flow control through nanoporous membrane

초록

Exosomes are considered promising biomarkers for liquid biopsy due to their capacity to transfer disease-related biomolecules between tissues or cells. However, their size range, spanning from 50 to 200 nm, presents a challenge in achieving efficient isolation. In this study, we propose a bidirectional flow control filtration (BFF) approach for exosome isolation. We employed a nanoporous polycarbonate membrane with 200 nm pores to eliminate apoptotic bodies, whereas a membrane featuring 50 nm pores was utilized to eliminate soluble proteins and concentrate exosomes. Backflush maneuvers were implemented during each phase to prevent the clogging of membrane pores, which could hinder the separation process. The BFF was applied to isolate exosomes from residual adipose tissue samples, achieving purity ratios 26 and 19 times higher compared to ultracentrifugation and direct-flow filtration, respectively, with a recovery rate of 68 %. In addition, the BFF achieved 4.4-fold higher purity and 1.9-fold higher throughput compared to size-exclusive chromatography for the isolation of exosomes from human serum. This method is scalable and facilitates a high processing rate of 1 L per hour, rendering it an optimal solution for generating large-scale exosomes for therapeutic applications. The method's scalability, minimal physical stress, and lack of chemical additives further enhance its suitability for therapeutic purposes.

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