2017
DOI: 10.1007/978-1-4939-7057-5_20
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Surface Profiling of Extracellular Vesicles from Plasma or Ascites Fluid Using DotScan Antibody Microarrays

Abstract: 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). Fl… Show more

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Cited by 6 publications
(6 citation statements)
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“…In contrast, Jamaly et al recently used various anticoagulants to isolate MV and measure their total plasma concentrations by NTA without finding any significant differences in numbers [68]. Apart from total MV numbers, the isolation of MV from serum drawn into lithium heparin tubes seems to result in the aggregation of platelet-derived MV, with sticky vesicle pellets and a reduction in platelet MV numbers, thereby specifically affecting certain MV subpopulations [69,70]. In addition to the anticoagulant, other preanalytical factors such as centrifugation, temperature, freeze-thaw cycles, or agitation can influence MV in blood samples, which should be taken into account when setting up isolation protocols for clinical studies [66,67,71].…”
Section: Analysis Of MV In Peripheral Bloodmentioning
confidence: 99%
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“…In contrast, Jamaly et al recently used various anticoagulants to isolate MV and measure their total plasma concentrations by NTA without finding any significant differences in numbers [68]. Apart from total MV numbers, the isolation of MV from serum drawn into lithium heparin tubes seems to result in the aggregation of platelet-derived MV, with sticky vesicle pellets and a reduction in platelet MV numbers, thereby specifically affecting certain MV subpopulations [69,70]. In addition to the anticoagulant, other preanalytical factors such as centrifugation, temperature, freeze-thaw cycles, or agitation can influence MV in blood samples, which should be taken into account when setting up isolation protocols for clinical studies [66,67,71].…”
Section: Analysis Of MV In Peripheral Bloodmentioning
confidence: 99%
“…Since PPP is often prepared by centrifugation at 2500-3000 × g, this step might result in a significant loss of larger MV, which already pellet at this force [9]. It has been proposed that two centrifugation steps at 1500× g might be preferable for MV isolation [70]; however, more elaborate sorting methods might be required to specifically isolate larger EV from blood.…”
Section: Analysis Of MV In Peripheral Bloodmentioning
confidence: 99%
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“…Later on, the same group reported the development of an array with capacity of screening 60 surface antigens from blood plasma without prior processing or enrichment [138]. Another established array, initially developed for leukaemia cells (DotScan), has been successfully implemented on vesicles for phenotypic analysis by fluorescence and chemiluminescence-based immunoassays [139].…”
Section: Microarraysmentioning
confidence: 99%
“…The current limit for reliable FAVS is ~100 nm (Momen-Heravi et al, 2012). An alternative would be magnetism activated vesicle sorting (MAVS), but this is not currently feasible either, because the size of the magnetic beads necessary for effective separation, which would have to be delivered into releasable vesicles to tag them, exceeds the diameter of the synaptic vesicle (Belov et al, 2016). To assess changes in the molecular composition of synaptic vesicles over time, I thus used a two-colour STED imaging approach on ultrathin slices.…”
Section: Experimental Approach To Determine Changes In Synaptic Vesic...mentioning
confidence: 99%