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Seminar: Special Guest Tingting Wang
June 18 @ 2:30 PM - 3:30 PM
Assessment of Extracellular Vesicles Purity and their Separation from Lipoproteins using Proteomics Toolbox
Speaker: Tingting Wang, Ph.D.
Institute for Bioscience and Biotechnology and National Institute of Standard and Technology (NIST)
Extracellular vesicles (EVs) are stable membrane structures that can deliver their cargo remotely and regulate fundamental cellular responses. A number of studies have demonstrated the pivotal role of EVs in intercellular communication of all multicellular organisms. Unfortunately, accurate analysis of EVs cargo faces a challenge of EVs isolation. Separation of EVs from non-EVs contaminants is poorly developed. For example, lipoprotein (LPs) particles abundant in human circulation have a similar size and density range and are typically co-isolated with EVs. Given that LPs have been also shown to carry extracellular microRNAs, the isolation protocols, which provide for separation of EVs and LPs, will help to discriminate EVs and non-EVs RNAs and unambiguously define the EVs functions. In the present study, we have capitalized on the fact that EVs membrane proteins presumably are the only membrane proteins in cell-free human fluids and quantitative analysis of these proteins by targeted mass spectrometry will allow evaluating the efficiency of EVs isolation.