International Society for Extracellular Vesicles

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Despite their small size, extracellular vesicles (EV) play important roles in normal physiological processes (e.g., immune response, neuronal function, and stem cell maintenance) and diseases (e.g., cancer and liver disease). A key bottleneck in EV research is the isolation of the vesicles, which has historically been accomplished via ultracentrifugation. However, ultracentrifugation is time-consuming, is not scalable, requires specialized equipment, may damage vesicles during the high-speed spins, and suffers from low yield. More recently, precipitation solutions have been utilized to simplify EV isolation protocols, but these techniques are often inconsistent, with low yield and reduced purity. Thus, there is a significant need for a method to rapidly isolate EVs without compromising purity or yield.

Here we describe the use of novel membranes conjugated to a proprietary, nonantibody-based EV-binding compound to selectively isolate EVs. The membranes, which we have named Capturem membranes, have been chemically modified to have increased surface area, which allows higher binding capacity while providing highly pure and concentrated samples. Additionally, the membranes have been assembled into benchtop centrifuge-compatible spin columns which can be used to isolate EVs in under 30 minutes.

Isolations performed with the Capturem EV isolation spin columns produced EVs of sizes comparable to experimental values reported in the literature; contained the key EV protein markers CD63, CD9, and Alix; and showed little or no expression of the EV-negative markers calnexin and albumin. Furthermore, we were able to collect ~3 ng total RNA from exosomes isolated from a single spin column (using an input of as little as 50 µl of plasma), enough total RNA to generate multiple cDNA libraries for downstream NGS analysis. These Capturem columns enable researchers studying EVs to accelerate the pace of their research by obtaining high yields of noncontaminated exosomes in a simple and rapid manner.

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Rapid, pure, and concentrated purification of extracellular vesicles

The Capturem Extracellular Vesicle Isolation Kit (Mini) consists of spinnable affinity columns containing novel, modified nylon membranes conjugated to a lectin-based, EV-binding compound, thus providing a unique solution for the isolation of concentrated, highly pure EVs. This system removes the bottleneck in EV research presented by traditional ultracentrifugation-based EV isolation methods, which suffer from low yield, reduced purity, and time-consuming protocols. With a room-temperature protocol that takes less than 30 minutes, the Capturem EV isolation kit can rapidly enrich EVs from plasma and other biofluids for downstream proteomic, genomic, and transcriptomic analyses.