title: Rapid isolation of extracellular vesicles from cell culture and biological fluids using a synthetic peptide with specific affinity for heat shock proteins.
abstract: 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.
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada; Department of Chemistry and Biochemistry, Université de Moncton, Moncton, New Brunswick, Canada.
authors.forename: Anirban
authors.initials: A
authors.lastname: Ghosh
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Michelle
authors.initials: M
authors.lastname: Davey
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Ian C
authors.initials: IC
authors.lastname: Chute
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Steven G
authors.initials: SG
authors.lastname: Griffiths
authors.affiliation: New England Peptide Inc., Gardner, Massachusetts, United States of America.
authors.forename: Scott
authors.initials: S
authors.lastname: Lewis
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Simi
authors.initials: S
authors.lastname: Chacko
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada; Department of Chemistry and Biochemistry, Université de Moncton, Moncton, New Brunswick, Canada; Department of Chemistry and Biochemistry, Mount Allison University, Sackville, New Brunswick, Canada.
authors.forename: David
authors.initials: D
authors.lastname: Barnett
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Crapoulet
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Fournier
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Andrew
authors.initials: A
authors.lastname: Joy
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Michelle C
authors.initials: MC
authors.lastname: Caissie
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Amanda D
authors.initials: AD
authors.lastname: Ferguson
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors.forename: Melissa
authors.initials: M
authors.lastname: Daigle
authors.affiliation: Department of Chemistry and Biochemistry, Mount Allison University, Sackville, New Brunswick, Canada.
authors.forename: M Vicki
authors.initials: MV
authors.lastname: Meli
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada; Department of Chemistry and Biochemistry, Université de Moncton, Moncton, New Brunswick, Canada; Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia, Canada; Department of Biology, University of New Brunswick, Saint John, New Brunswick, Canada.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Lewis
authors.affiliation: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada; Department of Chemistry and Biochemistry, Université de Moncton, Moncton, New Brunswick, Canada.
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Ouellette
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D006360:Heat-Shock Proteins / Q000378:metabolism*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D013058:Mass Spectrometry; D018625:Microscopy, Atomic Force; D008854:Microscopy, Electron; D010455:Peptides / Q000378:metabolism*; D040901:Proteomics; D022161:Transport Vesicles / Q000378:metabolism*; D014461:Ultracentrifugation
chemical_list: D006360:Heat-Shock Proteins; D010455:Peptides