EV-iTEC
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, inculding detemination of density by ....
- Tangential Flow Filtration (TFF)
- A scalable method ideal for isolating EVs from large volumes.
- Chromatography
- SEC: Gravity-based homemade Sepharose CL-4B columns and Izon columns with an automatic fraction collector are available for precise separation.
- ! NEW! ÄKTA Start sytsem available with CaptoCore
- 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.
- 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. CD63, CD81, CD29, and more).
- Western Blotting
- Confirm the presence of EV-specific proteins.
- Protein and Nucleic Acid Quantification
- MicroBCA and Qubit assays are available to measure EV content.
- Mass Spectrometry Sample Preparatio
- EV sample preparation and QC for proteomic analyses in collaboration with the Institute of Translational Proteomics & the associated Core Facility
