Molecular Analysis of Single Tumor-Derived Extracellular Vesicles with Improved Robustness and Accuracy
Year: 2026
Type: article
Abstract: Extracellular vesicles (EVs) have emerged as key analytes in liquid biopsy analysis, as they inherit the molecular characteristics of their parental cells. For cancer diagnosis and treatment monitoring, tumor-derived EVs can serve as surrogate markers of their originating tumor cells. However, technical challenges arise from EV heterogeneity, as molecular signals from a rare EV subpopulation are often neglected in bulk-EV analysis. This limitation underscores the importance of single-EV analysis, enabling the detection and analysis of individual tumor EVs. While multiple single-EV technologies have been introduced, accurately detecting and quantifying rare target EVs in clinical samples remains challenging in practice. Here, we developed a set of protocols that improve the selectivity and accuracy of single-EV analysis by minimizing false-positive signals and enhancing assay robustness. Specifically, we improved the validity of data processing for single-EV imaging, optimized EV labeling procedures, and applied colocalization analysis between cancer-specific markers and EV signals. We demonstrated that the approach is applicable to two orthogonal single-EV technologies, fluorescence imaging and nanoflow cytometry, yielding consistent results across the two methods and significantly improving the detection accuracy of tumor-derived EVs spiked in plasma samples. Collectively, we demonstrated precise single-EV profiling with high accuracy and selectivity, with evident benefits for the applicability of EV analysis in clinical studies.
Source: Analytical Chemistry
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7161962492