Construction of an Acetylcholinesterase Sensor Based on Synthesized Paramagnetic Nanoparticles, a Simple Tool for Neurotoxic Compounds Assay.
Magnetic particles (MPs) have been widely used in biological applications in recent years as a carrier for various molecules. Their big advantage is in repeated use of immobilized molecules including enzymes. Acetylcholinesterase (AChE) is an enzyme playing crucial role in neurotransmission and the enzyme is targeted by various molecules like Alzheimer's drugs, pesticides and warfare agents. In this work, an electrochemical biosensor having AChE immobilized onto MPs and stabilized through glutaraldehyde (GA) molecule was proposed for assay of the neurotoxic compounds. The prepared nanoparticles were modified by pure AChE and they were used for the measurement anti-Alzheimer's drug galantamine and carbamate pesticide carbofuran with limit of detection 1.5 µM and 20 nM, respectively. All measurements were carried out using screen-printed sensor with carbon working, silver reference, and carbon auxiliary electrode. Standard Ellman's assay was used for validation measurement of both inhibitors. Part of this work was the elimination of reversible inhibitors represented by galantamine from the active site of AChE. For this purpose, we used a lower pH to get the original activity of AChE after inhibition by galantamine. We also observed decarbamylation of the AChE-carbofuran adduct. Influence of organic solvents to AChE as well as repeatability of measurement with MPs with AChE was also established.
Faculty of Chemical Technology, University of Pardubice, Studentska 95, Pardubice CZ-53210, Czech Republic. st30827@student.upce.cz.|Faculty of Military Health Sciences, University of Defense, Trebesska 1575, Hradec Kralove CZ-50001, Czech Republic. st30827@student.upce.cz.
Adam
A
Kostelnik
Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, Brno CZ-61300, Czech Republic. pavel.kopel@mendelu.cz.|Central European Institute of Technology, Brno University of Technology, Purkynova 123, Brno CZ-61200, Czech Republic. pavel.kopel@mendelu.cz.
Pavel
P
Kopel
Faculty of Chemical Technology, University of Pardubice, Studentska 95, Pardubice CZ-53210, Czech Republic. Alexander.Cegan@upce.cz.
Alexander
A
Cegan
Faculty of Military Health Sciences, University of Defense, Trebesska 1575, Hradec Kralove CZ-50001, Czech Republic. miroslav.pohanka@gmail.com.|Department of Geology and Pedology, Faculty of Forestry and Wood Technology, Mendel University in Brno, Brno CZ-61300, Czech Republic. miroslav.pohanka@gmail.com.
Miroslav
M
Pohanka
D000110:Acetylcholinesterase; D015374:Biosensing Techniques; D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D053758:Nanoparticles; D009943:Organophosphorus Compounds; D010575:Pesticides
D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D009943:Organophosphorus Compounds; D010575:Pesticides; D000110:Acetylcholinesterase
acetylcholinesterase; carbofuran; electrochemistry; galantamine; magnetic particles; nanomaterial; nanoparticles; screen-printed sensor