Department of Chemistry, M.V. Lomonosov Moscow State University, Moscow 119991, Russia.
Chem Biol Interact. 2010 Sep 6;187(1-3):312-7. doi: 10.1016/j.cbi.2010.01.028. Epub 2010 Jan 25.
This paper reviews our previously published data and presents new results on biosensor assay of blood esterases. Tyrosinase and choline oxidase biosensors based on nanostructured polyelectrolyte films were developed for these purposes. Experiments were performed on the quantitative determination of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), carboxylesterase (CaE), and neuropathy target esterase (NTE) in samples of whole blood of rats, mice, and humans. Good agreement was found between biosensor and spectrophotometric assays for AChE, BChE, and CaE. No direct comparison could be made for NTE because its activity cannot be measured spectrophotometrically in whole blood. A new method of simultaneous quantitative determination of AChE and BChE in test mixtures is also described. This method represents a bifunctional biosensor for the simultaneous analysis of choline and phenol based on integration of individual sensors. Algorithms for calculation of separate concentrations of AChE and BChE in the mixture were developed. The mean error of calculated component concentrations was approximately 6% for binary test mixtures. The present work provides a foundation for building multiplexed systems for the simultaneous determination of multiple esterases with applications to biomonitoring for exposures to organophosphorus compounds.
本文回顾了我们之前发表的数据，并介绍了血液酯酶生物传感器分析的新结果。为此目的，开发了基于纳米结构聚电解质膜的酪氨酸酶和胆碱氧化酶生物传感器。在大鼠、小鼠和人全血样本中，进行了定量测定乙酰胆碱酯酶（AChE）、丁酰胆碱酯酶（BChE）、羧酸酯酶（CaE）和神经毒性靶酯酶（NTE）的实验。生物传感器和分光光度法在测定 AChE、BChE 和 CaE 方面具有良好的一致性。由于无法在全血中分光光度法测定 NTE 的活性，因此无法对其进行直接比较。还描述了一种在测试混合物中同时定量测定 AChE 和 BChE 的新方法。该方法代表了基于个体传感器集成的基于胆碱和苯酚的同时分析的双功能生物传感器。开发了用于计算混合物中单独 AChE 和 BChE 浓度的算法。计算得出的组分浓度的平均误差约为二元测试混合物的 6%。本工作为构建用于同时测定多种酯酶的多路复用系统奠定了基础，可应用于对有机磷化合物暴露的生物监测。
