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Development of characterization and fractionation/purification methods for organic nanoparticles, exemplified by extracellular vesicles from lactic bacteria.

Project


Project code: MRI-PBE-08-1012 EV von Milchprodukten
Contract period: 01.01.2019 - 30.06.2020
Purpose of research: Basic research
Keywords: Nutrition physiology

All cells release extracellular vesicles (EVs) delimited by a lipid bilayer membrane and which are considered as a way of communication between cells. Their relative stability as compared to synthetic liposomes, as well as the fact that they improved the stability and bioavailability of orally administered bioactive compounds such as curcumin make EVs interesting vehicles for delivery of bioactive compounds. Their future use as carrier of food ingredients and nutraceuticals requires further analyses. However, characterization of organic nanostructures is challenging. The aim of this project is to establish characterization methods for the analysis of organic nanoparticles using EVs from food-associated bacteria, as an example of future carrier of bioactive compounds. In this project we will: (i) Compare different raw purification methods (ultracentrifugation, kit-based) and further fractionation methods (Flow-field fractionation: AF4-MALS, density gradient ultracentrifugation, Size exclusion chromatography) by analyzing the protein profile (SDS-PAGE), size distribution (DLS) and Zeta-potential of obtained vesicle fractions from three bacterial species (Streptococcus salivarius ssp. thermophilus, Bifidobacterium bifidum, Lactobacillus delbrueckii ssp. bulgaricus). (ii) Test different visualization methods (electron microscopy: SEM/STEM, TEM with classical negative contrasting, embedment in methylcellulose or freeze-fracture approaches) and quantification methods (Flow-cytometry with fluorescent labelling).

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Subjects

Framework programme

BMEL - research cluster

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