Highly Efficient Isolation and Sensitive Detection of Small Extracellular Vesicles Using a Paper-Based Device

Abstract

Small extracellular vesicles (sEVs) play important roles in mediating intercellular communication and regulating biological processes. Facile sEV isolation is the essential and preliminary issue for their function investigation and downstream biomedical applications, while the traditional methods are challenged by tedious procedures, low purity, low yield, and potential damage. In this work, we developed an sEV isolation paper-based device (sEV-IsoPD) based on a three-dimensional (3D) paper chip, which is composed of a porous membrane for size exclusion and a metal–organic framework (MOF)/antibody-modified paper for immunoaffinity capture. In combination with a peristaltic pump-driven flow system, the sEV-IsoPD can efficiently isolate EV from cell culture medium and serum. Compared with the ultracentrifugation method, sEV-IsoPD exhibited a 5.1 times higher yield (1.7 × 109 mL–1), 1.6 times higher purity (1.6 × 1011 mg–1), and 7.5 times higher recovery (77.3%) with only 8.3% of the time (30 min) and 1.0% of the instrument cost ($710). Moreover, sEV concentration can be visually detected in a quantitative manner with this paper-based device with a linear range from 3.0 × 106 to 3.0 × 1010 mL–1 and a detection limit of 2.2 × 106 mL–1. The sEV-IsoPD provides an efficient and practical approach for the rapid isolation and visible detection of sEVs, which are promising for the preparation of sEVs and diagnosis of disease.

Recent research on material-based methods for isolation of extracellular vesicles

“…46 Later, Zhang et al continued to improve the paper-based capture method, and used metal–organic framework nanomaterial ZIF-90 to modify paper to immobilize a sufficient amount of CD63 antibodies, thereby constructing a three-dimensional device for efficient isolation of EVs. 47 In addition, considering that the number of fixed sites of antibodies is closely related to the recovery rate of EVs, graphene with abundant hydrophilic groups can attach a large number of specific antibodies. Fang et al functionalized CD63 antibodies to hydrophilic macroporous graphene foam.…”

Exploring the Evolution of Wuhu Iron Painting through Cultural and Tourism Integration in the Digital Age: An Innovative Development Perspective

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Label-Free Small Extracellular Vesicles Capturing Strategy for Lung Cancer Diagnosis and Typing Based on a Natural Polyphenol–Metal Three-Dimensional Network

“…Other strategies, such as affinity beads, microfluidic chips, and commercial precipitation kits, have also been developed for sEV isolation. − Most of these approaches are dependent on capture agents, such as antibodies and aptamers, which could recognize the biological molecules on the outer surface of sEVs. However, the quality and purity of the isolated sEVs may be compromised by the absence of surface antigens and the inactivation and degradation of these bioactive molecules, − which are not suitable for large-scale capture applications. Therefore, there is an unmet need to exploit a label-free, universal, low-cost, and easy scale-up approach for high-efficiency and high-purity isolation of sEVs.…”