Brief description

Aim:
To provide in-depth theoretical knowledge and hands-on experience in advanced methodologies for extracellular vesicle (EV) characterization, with a strong focus on antibody-based technologies, quantitative analysis, and critical evaluation of EV enrichment and recovery. The course aims to equip participants with practical and analytical skills needed to robustly characterize EV phenotype and assess EV isolation performance in research and clinical contexts.

Content:
Extracellular vesicles are key mediators of intercellular communication through the transfer of bioactive molecules such as proteins, lipids, and nucleic acids. While EVs play important roles in maintaining physiological homeostasis, they are also involved in disease development and progression, as both diseased cells and pathogens actively release EVs. These properties make EVs highly attractive as biomarkers for disease diagnosis, prognosis, and monitoring, as well as promising tools for therapeutic applications.

This course places a strong emphasis on advanced molecular and phenotypic characterization of EVs, with particular focus on antibody-based and high-sensitivity technologies used for EV analysis. Participants will explore how EV phenotype can be investigated using complementary approaches, ranging from broad biomarker screening to highly targeted analyses.

Key theoretical components include antibody–antigen interactions, molecular binding principles, and critical considerations related to specificity, sensitivity, and data interpretation in EV research. Particular attention is given to methodological challenges in EV enrichment, including sample type–specific considerations and sources of variability.

Practical training:
During the practical sessions, participants will gain hands-on experience in EV enrichment using size-exclusion chromatography (SEC) followed by advanced characterization using high-sensitivity flow cytometry (hFCM) and EV Array. In this course, hFCM will be used not only for EV phenotyping but also to estimate the recovery yield of EVs isolated by SEC, highlighting a quantitative and quality-control–oriented approach that is central to this course.

In addition, EV size distribution and particle concentration will be assessed using nanoparticle tracking analysis (NTA), enabling participants to critically compare different characterization platforms and understand their respective strengths and limitations.

Prerequisites

The students participating in this course should have basic knowledge of EVs and their nomenclature and biogenesis, although no previous experience with EV characterization is needed.

Contact

Associate Professor Maiken Mellergaard (maiken.pedersen@rn.dk)