Please use this identifier to cite or link to this item:
http://hdl.handle.net/2445/175540
Title: | Purification and analysis of exosomes |
Other Titles: | Purificació i anàlisi d’exosomes |
Author: | El Ouahabi Afettat, Oumaima |
Director/Tutor: | Benavente Moreno, Fernando J. (Julián) |
Keywords: | Electroforesi capil·lar Exosoma (vesícula) Dispersió dinàmica de llum Treballs de fi de grau Capillary electrophoresis Exosome (vesicle) Dynamic light scattering Bachelor's theses |
Issue Date: | Jan-2020 |
Abstract: | Exosomes are 30-150 nm extracellular nanovesicles that can be found in biological fluids such as blood, urine, sweat, tears, etc. They are secreted by all types of cells through exocytosis processes. Exosomes contain a wide variety of biological relevant molecules such as lipids, proteins, mRNAs, and microRNAs. They are thought to function as messengers between cells due to their ability to endocytosis and membrane fusion. Therefore, they are involved in many physiological and pathological processes, including tumor initiation and immune response. In this study, as an alternative to the techniques traditionally used to isolate exosomes, such as ultracentrifugation, it was explored polyethylene glycol (PEG) precipitation, which allows a facile, low-cost and effective isolation of exosomes from blood serum. Different variables of the method were studied, and the size of the isolated exosomes was measured by dynamic light scattering (DLS). In addition, a novel capillary electrophoresis (CE) with ultraviolet (UV) detection method was developed to complement these particle size measurements. Different concentrations of hydroxypropyl cellulose (HPC, 0.2, 0.5, 0.8% v/v) were added in the background electrolyte (BGE, 0.1 M tris + 0.25 M boric acid pH= 7.9) to reduce the adsorption of the exosomes to the inner wall of the separation capillary. The best results were obtained with a 0.5% v/v of HPC in the BGE. In order to homogenize the charge of the exosomes and make the separation only size dependent 0.1% v/v of sodium dodecyl sulfate (SDS) was also added to the BGE. Under these optimized conditions, a characteristic electrophoretic profile of the isolated exosomes was obtained, and separation showed the highest reproducibility and shortest analysis times |
Note: | Treballs Finals de Grau de Química, Facultat de Química, Universitat de Barcelona, Any: 2020, Tutor: Fernando Benavente |
URI: | http://hdl.handle.net/2445/175540 |
Appears in Collections: | Treballs Finals de Grau (TFG) - Química |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
TFG_QU El Ouahabi Afettat, Oumaima.pdf | 951.04 kB | Adobe PDF | View/Open |
This item is licensed under a Creative Commons License