EV - Core Facility
In recent years, extracellular vesicles (EVs) have attracted great interest in the scientific community due to their significant role in intercellular communication and mediating several physiological and pathological processes. However, the purification and characterization of the different EV types require, in addition to specific expertise, sophisticated isolation methods. Within the EV core facility, we offer these state-of-the-art techniques. In addition, we develop individual protocols to meet the different scientific questions. Besides, we offer services in training for various methods and study design consulting and can offer contacts to various downstream analyses in other core facilities based on optimized EV protocols.
Service Portfolio
Consultation Service:
- We offer expert guidance on designing EV-related experiments, ensuring compliance with MISEV (Minimal Information for Studies of Extracellular Vesicles) standards to enhance the robustness and reproducibility of your findings.
- Our support doesn’t end with your experiment. We provide post-experiment consultations to help you interpret data, troubleshoot issues, and ensure your results align with international standards and best practices for EV research.
EV Isolation Methods:
- Differential Ultracentrifugation (dUC)
High-speed separation using Beckman Coulter Optima XPN-80 and Optima Max-XP ultracentrifuges for EV recovery.
- Density Gradient Ultracentrifugation
Pure EV fractions can be isolated using iodixanol (OptiPrep™) or sucrose gradients.
- Tangential Flow Filtration (TFF)
A scalable method ideal for isolating EVs from large volumes.
- Size Exclusion Chromatography (SEC)
Gravity-based homemade Sepharose 2CLB columns and Izon columns with an automatic fraction collector are available for precise separation.
- Free-Flow Electrophoresis (FFE)
FFE is a powerful, cutting-edge technique used to separate extracellular vesicles based on their surface charge and electrophoretic mobility in a non-invasive manner. This method enables the enrichment of specific EV subpopulations while maintaining their structural integrity and functionality. It is particularly valuable for isolating EVs with subtle biochemical differences, such as variations in charge due to surface proteins or lipid composition.
EV characterization approaches:
- Nanoparticle Tracking Analysis (NTA)
Measure EV size, concentration, and zeta potential using a Zetaview.
- Nano-Flow Cytometry
Analyze EV surface markers using antibodies with fluorophores (e.g., Alexa488, FITC, APC). Several markers are available (e.g. CD9, CD63, CD81, CD29, and EpCam).
- Western Blotting
Confirm the presence of EV-specific proteins.
- RNA Isolation from EVs
Extract RNA for downstream analysis.
- Protein and Nucleic Acid Quantification
MicroBCA and Qubit assays are available to measure EV content.
- Mass Spectrometry Sample Preparation
EV sample preparation and QC for proteomic analyses in collaboration with the Institute of Translational Proteomics & the associated Core Facility
