High‐fidelity detection and sorting of nanoscale vesicles in viral disease and cancer

DOI Web Site Web Site 被引用文献2件
  • Aizea Morales‐Kastresana
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Thomas A. Musich
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Joshua A. Welsh
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • William Telford
    Experimental Immunology and Transplantation Branch National Cancer Institute, NIH Bethesda MD USA
  • Thorsten Demberg
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • James C. S. Wood
    Wake Forest School of Medicine Flow Cytometry Core Winston Salem NC USA
  • Marty Bigos
    Stanford University School of Medicine Stanford CA USA
  • Carley D. Ross
    Beckman Coulter Fort Collins CO USA
  • Aliaksander Kachynski
    Beckman Coulter Fort Collins CO USA
  • Alan Dean
    Beckman Coulter Fort Collins CO USA
  • Edward J. Felton
    Beth Israel Deaconess Medical Center Boston MA USA
  • Jonathan Van Dyke
    University of California, Davis Sacramento CA USA
  • John Tigges
    Beth Israel Deaconess Medical Center Boston MA USA
  • Vasilis Toxavidis
    Beth Israel Deaconess Medical Center Boston MA USA
  • David R. Parks
    Stanford University School of Medicine Stanford CA USA
  • W. Roy Overton
    QuantaCyte Corporation NJ USA
  • Aparna H. Kesarwala
    Radiation Oncology Branch National Cancer Institute, NIH Bethesda MD USA
  • Gordon J. Freeman
    Dana‐Farber Cancer Institute Boston MA USA
  • Ariel Rosner
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Stephen P. Perfetto
    Vaccine Research Center National Institute of Allergy and Infectious Disease, NIH Bethesda MD USA
  • Lise Pasquet
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Masaki Terabe
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Katherine McKinnon
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Veena Kapoor
    Experimental Immunology and Transplantation Branch National Cancer Institute, NIH Bethesda MD USA
  • Jane B. Trepel
    Developmental Therapeutics Branch National Cancer Institute, NIH Bethesda MD USA
  • Anu Puri
    Basic Research Lab National Cancer Institute, NIH Frederick MD USA
  • Hisataka Kobayashi
    Molecular Imaging Program National Cancer Institute, NIH Bethesda MD USA
  • Bryant Yung
    Theranostic Nanomedicine Section National Institute of Biomedical Imaging and Bioengineering, NIH Bethesda MD USA
  • Xiaoyuan Chen
    Theranostic Nanomedicine Section National Institute of Biomedical Imaging and Bioengineering, NIH Bethesda MD USA
  • Peter Guion
    Radiation Oncology Branch National Cancer Institute, NIH Bethesda MD USA
  • Peter Choyke
    Molecular Imaging Program National Cancer Institute, NIH Bethesda MD USA
  • Susan J. Knox
    Stanford University School of Medicine Stanford CA USA
  • Ionita Ghiran
    Beth Israel Deaconess Medical Center Boston MA USA
  • Marjorie Robert‐Guroff
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Jay A. Berzofsky
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA
  • Jennifer C. Jones
    Vaccine Branch National Cancer Institute National Institutes of Health (NIH) Bethesda MD USA

説明

<jats:title>ABSTRACT</jats:title><jats:p>Biological nanoparticles, including viruses and extracellular vesicles (EVs), are of interest to many fields of medicine as biomarkers and mediators of or treatments for disease. However, exosomes and small viruses fall below the detection limits of conventional flow cytometers due to the overlap of particle‐associated scattered light signals with the detection of background instrument noise from diffusely scattered light. To identify, sort, and study distinct subsets of EVs and other nanoparticles, as individual particles, we developed nanoscale Fluorescence Analysis and Cytometric Sorting (nanoFACS) methods to maximise information and material that can be obtained with high speed, high resolution flow cytometers. This nanoFACS method requires analysis of the instrument background noise (herein defined as the “reference noise”). With these methods, we demonstrate detection of tumour cell‐derived EVs with specific tumour antigens using both fluorescence and scattered light parameters. We further validated the performance of nanoFACS by sorting two distinct HIV strains to >95% purity and confirmed the viability (infectivity) and molecular specificity (specific cell tropism) of biological nanomaterials sorted with nanoFACS. This nanoFACS method provides a unique way to analyse and sort functional EV‐ and viral‐subsets with preservation of vesicular structure, surface protein specificity and RNA cargo activity.</jats:p>

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