Surface Profiling of Extracellular Vesicles from Plasma or Ascites Fluid Using DotScan Antibody Microarrays

Larissa Belov, Susannah Hallal, Kieran Matic, Jerry Zhou, Sandra Wissmueller, Nuzhat Ahmed, Sumaiya Tanjil, Stephen P. Mulligan, O. Giles Best, Richard J. Simpson, Richard I. Christopherson

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

6 Citations (Scopus)


DotScan antibody microarrays were initially developed for the extensive surface profiling of live leukemia and lymphoma cells. DotScan's diagnostic capability was validated with an extensive clinical trial using mononuclear cells from the blood or bone marrow of leukemia or lymphoma patients. DotScan has also been used for the profiling of surface proteins on peripheral blood mononuclear cells (PBMC) from patients with HIV, liver disease, and stable and progressive B-cell chronic lymphocytic leukemia (CLL). Fluorescence multiplexing allowed the simultaneous profiling of cancer cells and leukocytes from disaggregated colorectal and melanoma tumor biopsies after capture on DotScan. In this chapter, we have used DotScan for the surface profiling of extracellular vesicles (EV) recovered from conditioned growth medium of cancer cell lines and the blood of patients with CLL. The detection of captured EV was performed by enhanced chemiluminescence (ECL) using biotinylated antibodies that recognized antigens expressed on the surface of the EV subset of interest. DotScan was also used to profile EV from the blood of healthy individuals and the ascites fluid of ovarian cancer patients. DotScan binding patterns of EV from human plasma and other body fluids may yield diagnostic or prognostic signatures for monitoring the incidence, treatment, and progression of cancers.
Original languageEnglish
Title of host publicationSerum/Plasma Proteomics
Subtitle of host publicationMethods and protocols
EditorsDavid W. Greening, Richard J. Simpson
Place of PublicationSwitzerland
Number of pages39
EditionSecond edition
ISBN (Electronic)9781493970575
Publication statusPublished - 2017
Externally publishedYes

Publication series

NameMethods in molecular biology (Clifton, N.J.)
ISSN (Print)1064-3745


  • *Antibodies/immunology Antigens, CD *Ascites Biomarkers Cell Line *Extracellular Vesicles/metabolism Female Humans Leukemia, Lymphocytic, Chronic, B-Cell/blood Leukocytes, Mononuclear Luminescent Measurements/methods Ovarian Neoplasms/blood *Plasma/chemistry Protein Array Analysis/*methods *CD antigen *Chemiluminescence *Chronic lymphocytic leukemia *Exosomes *Ovarian cancer *Shed microvesicles
  • Shed microvesicles
  • Chronic lymphocytic leukemia
  • Chemiluminescence
  • Exosomes
  • CD antigen
  • Ovarian cancer


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