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Konstrukce cholinesterasoveho senzoru založeného na syntetizovaných magnetických částicích pro stanovení neurotoxických látek

机译:基于合成磁性颗粒的胆碱酯酶传感器的构建,用于测定神经毒性物质

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摘要

Magnetic particles (MPs) are 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 drugs, pesticides and warfare agents. In this work, an electrochemical biosensor having AChE immobilized onto MPs and stabilized through glutaraldehyde 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 drug galantamine and carbamate pesticide carbofuran. 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 elimination of reversible inhibitors represented by the galantamine from active site of AChE. For this purpose, we used lower pH to get original activity of AChE after inhibition by galantamine. We also observed decarbamylation of AChE-carbofuran adduct. Influence of organic solvents to AChE as well as repeatability of measurement with MPs with AChE was also established.
机译:近年来,磁性颗粒(MP)作为各种分子的载体广泛用于生物学应用。它们的最大优点是可以重复使用包括酶在内的固定分子。乙酰胆碱酯酶(AChE)是一种在神经传递中起关键作用的酶,该酶被各种分子靶向,例如阿尔茨海默氏病药物,杀虫剂和战剂。在这项工作中,提出了一种将AChE固定在MP上并通过戊二醛分子稳定化的电化学生物传感器,用于分析神经毒性化合物。所制得的纳米颗粒经纯AChE修饰,可用于测定抗阿尔茨海默病药物加兰他敏和氨基甲酸酯农药呋喃丹。所有测量均使用带有碳工作,银参比和碳辅助电极的丝网印刷传感器进行。标准Ellman分析法用于两种抑制剂的验证测量。这项工作的一部分是从AChE活性位点消除以加兰他敏为代表的可逆抑制剂。为此,我们用较低的pH值获得了加兰他敏抑制后的AChE原始活性。我们还观察到了AChE-呋喃呋喃加合物的脱氨基甲酰化作用。还确定了有机溶剂对AChE的影响以及用AChE进行MP测量的可重复性。

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