Development and comprehensive comparison of two on-line capillary electrophoretic methods for beta-secretase inhibitor screening

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F17%3A00095133" target="_blank" >RIV/00216224:14310/17:00095133 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1016/j.chroma.2017.08.065" target="_blank" >https://doi.org/10.1016/j.chroma.2017.08.065</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.chroma.2017.08.065" target="_blank" >10.1016/j.chroma.2017.08.065</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Development and comprehensive comparison of two on-line capillary electrophoretic methods for beta-secretase inhibitor screening

Popis výsledku v původním jazyce

Alzheimer’s disease is the most common cause of dementia, afflicting over 34 million patients worldwide. Since beta-secretase is a rate-limiting enzyme of the production of neurotoxic beta-amyloid peptide oligomers abnormally accumulated in the affected brain tissue, its specific inhibition appears to be a promising approach to slowing down or even stopping the progression of the disease. Hence two on-line capillary electrophoretic methods for studies of beta-secretase activity based on the principles of transverse diffusion of laminar flow profiles and electrophoretically mediated microanalysis were developed, both using a simple unlabeled peptide substrate and UV detection. The optimized procedures were thoroughly validated and applied for determining the enzyme’s kinetic parameters and the inhibition characteristics of two potent probe inhibitors. The resulting values were found to be comparable to literature data obtained with other analytical techniques. The suitability of the employed methodologies for different experimental designs is discussed on the basis of a statistical evaluation of the experimental data. The presented methods constitute a miniaturized and fully automated tool, which should be suitable for kinetic and inhibition studies of beta-secretase as a target for Alzheimer’s disease drug discovery in the early stages of the development of a new drug.

Název v anglickém jazyce

Development and comprehensive comparison of two on-line capillary electrophoretic methods for beta-secretase inhibitor screening

Popis výsledku anglicky

Alzheimer’s disease is the most common cause of dementia, afflicting over 34 million patients worldwide. Since beta-secretase is a rate-limiting enzyme of the production of neurotoxic beta-amyloid peptide oligomers abnormally accumulated in the affected brain tissue, its specific inhibition appears to be a promising approach to slowing down or even stopping the progression of the disease. Hence two on-line capillary electrophoretic methods for studies of beta-secretase activity based on the principles of transverse diffusion of laminar flow profiles and electrophoretically mediated microanalysis were developed, both using a simple unlabeled peptide substrate and UV detection. The optimized procedures were thoroughly validated and applied for determining the enzyme’s kinetic parameters and the inhibition characteristics of two potent probe inhibitors. The resulting values were found to be comparable to literature data obtained with other analytical techniques. The suitability of the employed methodologies for different experimental designs is discussed on the basis of a statistical evaluation of the experimental data. The presented methods constitute a miniaturized and fully automated tool, which should be suitable for kinetic and inhibition studies of beta-secretase as a target for Alzheimer’s disease drug discovery in the early stages of the development of a new drug.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10600 - Biological sciences

Projekt

<a href="/cs/project/GA16-06106S" target="_blank" >GA16-06106S: Vysoce efektivní systém založený na kapilární elektroforéze pro screening inhibitorů beta-sekretázy jako terapeutického cíle pro Alzheimerovu chorobu</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Chromatography A

ISSN

0021-9673

e-ISSN

1873-3778

Svazek periodika

1518

Číslo periodika v rámci svazku

October

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

8

Strana od-do

89-96

Kód UT WoS článku

000412621200010

EID výsledku v databázi Scopus

2-s2.0-85028615141