Rapid and efficient isolation platform for plasma extracellular vesicles: EV‐FISHER

Abstract

Extracellular vesicles (EVs) have found diverse applications in clinical theranostics. However, the current techniques to isolate plasma EVs suffer from burdensome procedures and limited yield. Herein, we report a rapid and efficient EV isolation platform, namely, EV-FISHER, constructed from the metal-organic framework featuring cleavable lipid probes (PO 4 3− -spacer-DNA-cholesterol, PSDC). The EV-FISHER baits EVs from plasma by cholesterol and separates them with an ordinary centrifuge. The captured EVs could be released and collected upon subsequent cleavage of PSDC by deoxyribonuclease I. We conclude that EV-FISHER dramatically outperforms the ultracentrifugation (UC) in terms of time (∼40 min vs. 240 min), isolation efficiency (74.2% vs. 18.1%), and isolation requirement (12,800 g vs. 135,000 g). In addition to the stable performance in plasma, EV-FISHER also exhibited excellent compatibility with downstream single-EV flow cytometry, enabling the identification of glypican-1 (GPC-1) EVs for early diagnosis, clinical stages differentiation, and therapeutic efficacy evaluation in breast cancer cohorts. This work portrays an efficient strategy to isolate EVs from complicated biological fluids with promising potential to facilitate EVs-based theranostics.

Bacterial extracellular vesicles: A position paper by the microbial vesicles task force of the Chinese society for extracellular vesicles

“…This new diagnostic approach offers insights into the diagnosis of BEVs 64 . In recent years, new technologies, such as EV‐FISHER, 65 aptamer‐based methods, 66 and droplet‐based single‐exosome‐counting, 67 have enabled the development of diagnostic BEVs. Previous study on diagnostic BEVs …”

Profiling extracellular vesicle surface proteins with 10 µL peripheral plasma within 4 h

“…EVs carry a variety of biomolecules inherited from their parent cells (including nucleic acids, proteins, lipids and metabolites), and are involved in a wide range of physiological and pathological processes (Mathieu et al., 2019). Notably, many tumour‐derived EV proteins have been identified as important biomarkers for early cancer diagnosis (Pan et al., 2022; Qian et al., 2022; Xu et al., 2018), where the type and expression levels of EV proteins can indicate the cancer classification and progression status (van Niel et al., 2018). However, the small clinical sample volumes, confounded with interference by non‐EV components and the normally low expression levels of target proteins pose great challenges for specific EV isolation and protein analysis (Yu et al., 2022), both of which are necessary for an accurate clinical diagnosis.…”

“…as important biomarkers for early cancer diagnosis (Pan et al, 2022;Qian et al, 2022;Xu et al, 2018), where the type and expression levels of EV proteins can indicate the cancer classification and progression status (van Niel et al, 2018). However, the small clinical sample volumes, confounded with interference by non-EV components and the normally low expression levels of target proteins pose great challenges for specific EV isolation and protein analysis (Yu et al, 2022), both of which are necessary for an accurate clinical diagnosis.…”

Challenges and Opportunities for Extracellular Vesicles in Clinical Oncology Therapy

“…In recent years, EVs have been found in a variety of physiological fluids including blood, urine, saliva, breast milk, amniotic fluid, ascites, cerebrospinal fluid, bile, and semen [ 8 , 9 , 10 , 11 , 12 ]. More importantly, EVs can be directly isolated from physiological fluid in solid organs including the liver, heart, brain, and others [ 13 , 14 , 15 , 16 , 17 ]. Originally, the majority of studies focused on clinical diagnosis; however, in recent years, EVs have shown a bright future as an excellent therapeutic vehicle for treating diseases, especially cancer.…”