Acetylcholinesterase-ISFET based system for the detection of acetylcholine and acetylcholinesterase inhibitors

Biosensors & Bioelectronics
Aviad HaiShlomo Yitzchaik

Abstract

A bioelectronic hybrid system for the detection of acetylcholine esterase (AChE) catalytic activity was assembled by way of immobilizing the enzyme to the gate surface of an ion-sensitive field-effect transistor (ISFET). Photometric methods used to characterize bonded enzyme and linker layers on silicon substrates confirm the existence of a stable amino-cyanurate containing AChE monolayer. The transduction of the enzyme-functionalized ISFET, in ionic solutions, is detected in response to application of acetylcholine (ACh). Recorded sensitivity of the modified ISFET to ACh has reached levels of up to 10(-5)M. The Michaelis-Menten constant of the immobilized AChE is only moderately altered. Nevertheless, the maximum reaction velocity is reduced by over an order of magnitude. The ISFET response time to bath or ionophoretic application of ACh from a micropipette was in the range of a second. The catalytic activity of the immobilized AChE is inhibited in a reversible manner by eserine, a competitive inhibitor of AChE. We conclude that the immobilized enzyme maintains its pharmacological properties, and thus the described bioelectronic hybrid can serve as a detector for reagents that inhibit AChE activity.

References

Jan 16, 2003·Journal of the American Chemical Society·Vered Pardo-YissarItamar Willner

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Citations

Nov 19, 2010·Bioanalysis·Sonia CentiMarco Mascini
Mar 20, 2012·Journal of Neuroscience Methods·Michael GebinogaAndreas Schober
Mar 11, 2009·IEEE Transactions on Bio-medical Engineering·Tung-Ming Pan, Kao-Ming Liao
Apr 3, 2008·Biochimica Et Biophysica Acta·Avraham BrombergGad Frishman
Jul 13, 2016·Journal of Enzyme Inhibition and Medicinal Chemistry·Šárka Štěpánková, Katarína Vorčáková
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Sep 17, 2009·Electrophoresis·Aiping Schuchert-Shi, Peter C Hauser
Jan 30, 2018·Luminescence : the Journal of Biological and Chemical Luminescence·Alsu MukhametshinaOleg Sinyashin
Jun 4, 2011·Small·Matthew S Makowski, Albena Ivanisevic
Aug 2, 2017·Engineering in Life Sciences·Bhuvanagayathri RajangamAlbert Ivanov Krastanov
Mar 19, 2009·Journal of the American Chemical Society·Ilya GoykhmanShlomo Yitzchaik
Aug 12, 2008·Langmuir : the ACS Journal of Surfaces and Colloids·Andreas HärtlJose A Garrido
May 2, 2013·Langmuir : the ACS Journal of Surfaces and Colloids·Keren YaakobiHanna Rapaport
Sep 27, 2018·Langmuir : the ACS Journal of Surfaces and Colloids·Shlomo YitzchaikRoie Yerushalmi

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