Abstract

Luncheon Technology Spotlight: Next Generation Affinity-based Extracellular Vesicle Isolation Technologies that Rely on a Synthetic Peptide or Hyaluronic Acid

Anirban Ghosh, Research Scientist, Atlantic Cancer Research Institute

Given the growing evidence that circulating extracellular vesicles (EVs) have tremendous potential for minimally-invasive diagnostics and therapeutic applications, there is a great demand for simple, efficient and clinically-compatible affinity-based EV isolation methods for use with biofluids. Currently available polymer-based EV isolation methods facilitate non-specific precipitation of other materials with EVs and the polymers themselves are toxic for therapeutic use. Most affinity-based EV-isolation methods rely on antibodies directed against EV surface marker(s) which can be costly and difficult to scale-up. We have developed and validated robust affinity-based EV-capture technologies that use (i) a synthetic peptide (Vn96) and (ii) hyaluronic acid (HA). The Vn96 peptide targets canonical heat shock proteins that are present on the surface of EVs and we discovered the propensity for long-chain HA to bind to EVs and thereby facilitate their isolation. We have examined the efficacy of these methods for downstream analyses, clinical compatibility, liquid biopsy assays (biomarker and mutation detection) and platform-versatility using cell-culture conditioned media and human body fluids as sources of EVs. Our results indicate that the scalability, quality, platform versatility and cost-effectiveness of our affinity-based EV isolation technologies provide multiple advantages over currently-available affinity-capture methods for EV isolation.