Reference : Challenges in the determination of amyloid oligomeric species by two electrophoretic ...
Scientific congresses and symposiums : Poster
Human health sciences : Pharmacy, pharmacology & toxicology
http://hdl.handle.net/2268/214727
Challenges in the determination of amyloid oligomeric species by two electrophoretic techniques
English
Napp, Aurore mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
Houbart, Virginie mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
Demelenne, Alice mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
Servais, Anne-Catherine mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
Fillet, Marianne mailto [Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments >]
18-Sep-2017
No
International
CE-PHARM 2017
du 17 septembre 2017 au 20 septembre 2017
CASSS
Boston
USA
[en] alpha-synuclein ; Derivatization ; Capillary gel electrophoresis ; SDS-PAGE
[en] Parkinson’s disease is a frequent degenerative disorder, and for the moment the diagnosis is mainly clinical. When the first symptoms appear, loss of more than 70% of the dopaminergic cells already occurred. Knowing that, it is of high interest to have one (or more) reliable biomarker(s) at our disposal to diagnose Parkinson before the first symptoms appear. Alpha-synuclein (aSyn) is a protein physiologically expressed at high level by neuronal cells, under a monomeric form. This protein would play a critical role in the development of the disease because under certain conditions, aSyn is capable of self-assembly to form fibrils like those found in Lewy bodies. Other intermediate soluble forms like dimers and oligomers are also formed. As these forms seems to be the toxic species, they are the center of many attentions. The quantification of each form would be a great help, but for the moment only the total forms (of monomeric or oligomeric) can be quantified. In this study, aSyn oligomers were generated after optimization of incubation conditions (pH, temperature, agitation, …). Then, different approaches were investigated to detect and follow the different species formed during the aggregation. We analyzed the oligomers by capillary gel electrophoresis (CGE) and SDS-PAGE. We found that capillary gel electrophoresis is a promising automated technique to analyze aSyn oligomers, due to the fact that it separates the aggregates according to their size, like the SDS-PAGE, but with more advantages.
To gain sensitivity and selectivity by CGE, we used a laser-induced fluorescence detector. As aSyn do not have a native fluorescence, we derivatized it. After careful screening and optimization of various derivatization reagents, we could quantify with high sensitivity aSyn oligomers by CGE-LIF. We realized different calibration curves, and we had promising results that will allow us to quantify the different aSyn oligomeric forms in biological fluids.
Centre Interdisciplinaire de Recherche sur le Médicament - CIRM
Researchers ; Professionals
http://hdl.handle.net/2268/214727

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