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Browsing by Subject "ultrafiltraatio"

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  • Niemeläinen, Elina (2020)
    Extracellular vesicles (EVs) are lipid bilayer-enclosed nano-sized particles that are found in all body fluids. EVs are part of normal cell functions and they can carry for example proteins, RNA and lipids between cells. This makes them potential candidates as drug delivery vehicles. When nanoparticles are introduced to blood plasma, a plasma protein structure is formed on their surface, called the protein corona. The formation of a protein corona is a dynamic process, and the proteins are binding to the corona depending on their affinity to the nanoparticle surface. When administrating nanoparticles to cells, protein corona has a big impact on the half-life and cellular uptake of the particles. The aim of this work was to study the plasma protein corona of PC-3 derived EVs, and to investigate methods for protein corona isolation. First, EVs and lipoprotein particles were removed from fresh frozen plasma by membrane filtration. Using this filtrated plasma, we then tested plasma-EV incubation and removing of free plasma proteins by simple ultrafiltration. Removing of free plasma proteins was also examined by size-exclusion chromatography (SEC). The resulting EV-protein corona complex was visualized with SDS-PAGE. EVs were characterized with nanoparticle tracking analysis and western blotting. Of these two methods used, SEC appeared to be more convenient and efficient way to remove the free plasma proteins from the EV-protein corona complex. The method still requires further development and testing in order to perform optimally.