Rapid Isolation of Extracellular Vesicles from Cell Culture and Biological Fluids Using a Synthetic Peptide with Specific Affinity for Heat Shock Proteins

• DOI: 10.1371/journal.pone.0110443
• Publication Date: 2014-10-20T17:50:23Z
• Authors: Anirban Ghosh; Michelle Davey; Ian C. Chute; Steven G. Griffiths; Scott Lewis; Simi Chacko; David Barnett; Nicolas Crapoulet; Sébastien Fournier; Andrew Joy; Michelle C. Caissie; Amanda D. Ferguson; Melissa Daigle; M. Vicki Meli; Stephen M. Lewis; Rodney J. Ouellette
• Keywords: Proteomics; 0303 health sciences; Science; Q; Blotting, Western; R; High-Throughput Nucleotide Sequencing; Microscopy, Atomic Force; Mass Spectrometry; Microscopy, Electron; 03 medical and health sciences; Cell Line, Tumor; Medicine; Humans; Peptides; Transport Vesicles; Ultracentrifugation; Heat-Shock Proteins; Research Article

Recent studies indicate that extracellular vesicles are an important source material for many clinical applications, including minimally-invasive disease diagnosis. However, challenges for rapid and simple extracellular vesicle collection have hindered their application. We have developed and validated a novel class of peptides (which we named venceremin, or Vn) that exhibit nucleotide-independent specific affinity for canonical heat shock proteins. The Vn peptides were validated to specifically and efficiently capture HSP-containing extracellular vesicles from cell culture growth media, plasma, and urine by electron microscopy, atomic force microscopy, sequencing of nucleic acid cargo, proteomic profiling, immunoblotting, and nanoparticle tracking analysis. All of these analyses confirmed the material captured by the Vn peptides was comparable to those purified by the standard ultracentrifugation method. We show that the Vn peptides are a useful tool for the rapid isolation of extracellular vesicles using standard laboratory equipment. Moreover, the Vn peptides are adaptable to diverse platforms and therefore represent an excellent solution to the challenge of extracellular vesicle isolation for research and clinical applications.