About the Doctoral dissertation

Extracellular vesicles (EVs) are membrane-bound nanoparticles re-leased by all cells, carrying cargo that reflects their origin and influ-ences distant cells. Because of this, they are promising biomarkers and therapeutic targets.

In 2011, EVs were mainly analyzed by immunoblotting or flow cytom-etry, requiring large protein amounts (10–30 mg) and extensive isola-tion. In 2013, we introduced protein microarrays to capture and char-acterize EVs from just 10 µL of plasma or cell culture supernatant. This method allows simultaneous, rapid, and sensitive detection of many proteins with minimal sample use. By 2015, the platform was ex-panded to 60 antigens with higher sensitivity and throughput.

To move toward clinical use, the glass-slide arrays were replaced by a 96-well plate format, enabling easier handling in research and diag-nostic labs. However, pre-analytical factors—such as blood collection tubes, storage, and handling—were shown to affect outcomes, em-phasizing the need for standardized protocols.

Venous blood is the standard source of EVs, but we demonstrated that intact EVs can also be recovered from dried blood spots (DBS). DBS samples can be collected outside clinical settings and mailed, making EV-based screening more feasible.

Recent advances include dual-color detection for simultaneous bi-omarker measurement and integration with Single Molecule Localiza-tion Microscopy (SMLM), enabling highly sensitive marker quantifica-tion and spatial localization of EVs by surface markers, cargo, size, and shape.