%0 Journal Article
%T Detection of Extracellular Vesicle Surface Proteins and microRNAs in Clustered Vesicles by Flow Cytometry Using a Single Step in Situ Approach
%A Kim Se-En
%A Mi Jeong Kim
%J Bulletin of Pioneering Researches of Medical and Clinical Science
%@ 3006-2659
%D 2021
%V 1
%N 1
%R 10.51847/uzNTCM0gZ0
%P 57-68
%X Given the heterogeneous nature of cancers, accurate and specific diagnosis necessitates multiplexed biomarker detection. Extracellular vesicles (EVs) have emerged as promising candidates for liquid biopsy biomarkers in cancer due to their molecular cargo. Nevertheless, their nanoscale size poses significant technical challenges, highlighting the need for an efficient and straightforward detection strategy. In this study, we established a single step in situ detection approach capable of simultaneously identifying both surface proteins and internal miRNAs of EVs through flow cytometry. To achieve multiplexed detection in normal and cancer-derived EVs, we employed a CD63 antibody alongside molecular beacon-21. Moreover, a phospholipid–polymer–phospholipid conjugate was utilized to promote EV clustering, as confirmed by nanoparticle tracking analysis, thereby amplifying the detection signal. This method successfully enabled the differentiation of cancer-derived EVs using a flow cytometer. Overall, our findings demonstrate that single step in situ detection of multiple EV biomarkers via flow cytometry represents a rapid, simple, and minimally invasive liquid biopsy technique with potential applications in cancer and other disease diagnostics.
%U https://bprmcs.com/article/detection-of-extracellular-vesicle-surface-proteins-and-micrornas-in-clustered-vesicles-by-flow-cyto-gcjjys4y1vmstkq