Multi-Surface Antigen Staining of Larger Extracellular Vesicles
Larger extracellular vesicles, microparticles (MPs) or microvesicles (MVs), especially their acquisition and characterization by flow cytometry (FACS), is increasingly in focus of clinical/translational research efforts. Several laboratories have shown that MPs/MVs might be suitable for the diagnosis and predicting prognosis in various diseases including cancer. However, FACS staining of larger extracellular vesicles (EVs) can be difficult and results potentially in false positive and inconsistent data interpretation. Despite that FACS equipment is well maintained and the operators have ample experience, a reliable and for larger EVs optimized staining protocol is missing. Here, we aim to close that gap and provide a working multi-antibody staining protocol for larger EVs isolated from human serum samples. We describe in detail the needed steps as currently done in our laboratory. Staining is demonstrated exemplarily for multi-antibody mix including CD147, a potential cancer marker if applied in combination with other MP/MV surface markers.
Key wordsLarger extracellular vesicles Microparticles Microvesicles Surface antigen staining Antibody Flow cytometry
This work was supported by a Deutsche Krebshilfe grant (111184) to Miroslaw Kornek; and by the Alexander von Humboldt Foundation, Sofja Kovalevskaja Award to Veronika Lukacs-Kornek.
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