Surface Profiling of Extracellular Vesicles Using DotScan Antibody Microarrays

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.

University of Sydney researchers 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.


Schematic representation of capture and detection of cells and EV on immobilized antibody dots of DotScan: (a) Captured cells are usually detected by optical scanning; however, fluorescent antibodies can be used for fluorescence multiplexing of cells (b) EV are detected with fluorescent antibodies (b), or (c) by enhanced chemiluminescence (ECL), with biotinylated antibodies and Streptavidin Poly-HRP

Belov L, Hallal S, Matic K, Zhou J, Wissmueller S, Ahmed N, Tanjil S, Mulligan SP, Best OG, Simpson RJ, Christopherson R. (2017) Surface Profiling of Extracellular Vesicles from Plasma or Ascites Fluid Using DotScan Antibody Microarrays. Methods Mol Biol 1619:263-301. [abstract]

Leave a Reply

Your email address will not be published. Required fields are marked *