Capillary electrophoresis-integrated immobilized enzyme reactor for kinetic and inhibition assays of beta-secretase as the Alzheimer's disease drug target

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F19%3A00107212" target="_blank" >RIV/00216224:14310/19:00107212 - isvavai.cz</a>

Výsledek na webu

<a href="https://onlinelibrary.wiley.com/doi/full/10.1002/jssc.201800947" target="_blank" >https://onlinelibrary.wiley.com/doi/full/10.1002/jssc.201800947</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/jssc.201800947" target="_blank" >10.1002/jssc.201800947</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Capillary electrophoresis-integrated immobilized enzyme reactor for kinetic and inhibition assays of beta-secretase as the Alzheimer's disease drug target

Popis výsledku v původním jazyce

Capillary electrophoresis-integrated immobilized enzyme reactors are becoming an increasingly popular alternative for enzyme kinetic and inhibition assays thanks to their unique set of features including cost effectiveness, repeated use of the enzyme, minuscule sample consumption, rapid analysis time and easy automation. In this work we present the development and application of a CE-integrated immobilized enzyme reactor based on magnetic particles for kinetic and inhibition studies of beta secretase, a key enzyme in the development of Alzheimer’s disease and a promising drug target. We document the optimization of the immobilization procedure, characterization of immobilized beta secretase, optimization of a mutually compatible incubation protocol and separation method as well as the production of the CE-integrated immobilized enzyme reactor. The applicability of the CE-integrated immobilized enzyme reactor was demonstrated by kinetic assay with an unlabelled substrate and by inhibition assays using three structurally different reference inhibitors. The resulting kinetic and inhibition parameters clearly support the applicability of the herein presented method as well as document the fundamental phenomena which need to be taken in account when comparing the results to other methods.

Název v anglickém jazyce

Capillary electrophoresis-integrated immobilized enzyme reactor for kinetic and inhibition assays of beta-secretase as the Alzheimer's disease drug target

Popis výsledku anglicky

Capillary electrophoresis-integrated immobilized enzyme reactors are becoming an increasingly popular alternative for enzyme kinetic and inhibition assays thanks to their unique set of features including cost effectiveness, repeated use of the enzyme, minuscule sample consumption, rapid analysis time and easy automation. In this work we present the development and application of a CE-integrated immobilized enzyme reactor based on magnetic particles for kinetic and inhibition studies of beta secretase, a key enzyme in the development of Alzheimer’s disease and a promising drug target. We document the optimization of the immobilization procedure, characterization of immobilized beta secretase, optimization of a mutually compatible incubation protocol and separation method as well as the production of the CE-integrated immobilized enzyme reactor. The applicability of the CE-integrated immobilized enzyme reactor was demonstrated by kinetic assay with an unlabelled substrate and by inhibition assays using three structurally different reference inhibitors. The resulting kinetic and inhibition parameters clearly support the applicability of the herein presented method as well as document the fundamental phenomena which need to be taken in account when comparing the results to other methods.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

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

2019

Kód důvěrnosti údajů

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

Název periodika

Journal of Separation Science

ISSN

1615-9306

e-ISSN

1615-9314

Svazek periodika

42

Číslo periodika v rámci svazku

5

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

10

Strana od-do

1067-1076

Kód UT WoS článku

000461674900013

EID výsledku v databázi Scopus

2-s2.0-85061060350