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@article{1480516,
author = {Schejbal, Jan and Šefraná, Šárka and Řemínek, Roman and Glatz, Zdeněk},
article_location = {Weinheim},
article_number = {5},
doi = {http://dx.doi.org/10.1002/jssc.201800947},
keywords = {beta-secretase; capillary electrophoresis; immobilized enzyme reactor; inhibition; magnetic particles},
language = {eng},
issn = {1615-9306},
journal = {Journal of Separation Science},
title = {Capillary electrophoresis-integrated immobilized enzyme reactor for kinetic and inhibition assays of beta-secretase as the Alzheimer's disease drug target},
url = {https://onlinelibrary.wiley.com/doi/full/10.1002/jssc.201800947},
volume = {42},
year = {2019}
}
TY - JOUR
ID - 1480516
AU - Schejbal, Jan - Šefraná, Šárka - Řemínek, Roman - Glatz, Zdeněk
PY - 2019
TI - Capillary electrophoresis-integrated immobilized enzyme reactor for kinetic and inhibition assays of beta-secretase as the Alzheimer's disease drug target
JF - Journal of Separation Science
VL - 42
IS - 5
SP - 1067-1076
EP - 1067-1076
PB - WILEY-VCH Verlag GmbH & Co.
SN - 16159306
KW - beta-secretase
KW - capillary electrophoresis
KW - immobilized enzyme reactor
KW - inhibition
KW - magnetic particles
UR - https://onlinelibrary.wiley.com/doi/full/10.1002/jssc.201800947
L2 - https://onlinelibrary.wiley.com/doi/full/10.1002/jssc.201800947
N2 - 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.
ER -
SCHEJBAL, Jan, Šárka ŠEFRANÁ, Roman ŘEMÍNEK a Zdeněk GLATZ. Capillary electrophoresis-integrated immobilized enzyme reactor for kinetic and inhibition assays of beta-secretase as the Alzheimer's disease drug target. \textit{Journal of Separation Science}. Weinheim: WILEY-VCH Verlag GmbH \&{} Co., 2019, roč.~42, č.~5, s.~1067-1076. ISSN~1615-9306. Dostupné z: https://dx.doi.org/10.1002/jssc.201800947.
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