To acquire high-quality EVs from complex samples like plasma, a simplified dichotomic SEC method was established. Utilizing the Flow NanoAnalyzer, particle concentration and size distribution was first determined. The purity was assessed by implementing CFSE labelling of EVs. The dichotomic SEC method was found to separate particles in serum or plasma into two components: EVs and non-vesicular contaminates. Corroborating this, proteomics showed that the dichotomic SEC method can remove different co-isolating contaminant proteins from human plasma with comparable purity to that of UC. Collectively, this isolation method shows an intriguing potential in the preparation of EVs toward clinical testing and/or basic research.
Figure 1. Optimization of SEC for plasma EV isolation
By implementing the multi-analysis capabilities of the Flow NanoAnalyzer it was found that the dichotomic SEC and ultracentrifugation could isolate EVs from human plasma with comparable purity and even higher extraction rate by SEC.
J Extracell Vesicles, 2021, 10:e12145.