Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering, Faculty of Biology, Environmental Sciences and Chemistry, University of Alcalá, E-28871, Alcalá de Henares, Madrid, Spain.
Anal Chem. 2015;87(10):5074-8. doi: 10.1021/acs.analchem.5b00979. Epub 2015 May 2.
An electrochemical microfluidic strategy for the separation and enantiomeric detection of D-methionine (D-Met) and D-leucine (D-Leu) is presented. These D-amino acids (D-AAs) act as biomarkers involved in relevant diseases caused by Vibrio cholerae. On a single layout microfluidic chip (MC), highly compatible with extremely low biological sample consumption, the strategy allowed the controlled microfluidic D-AA separation and the specific reaction between D-amino acid oxidase (DAAO) and each D-AA biomarker avoiding the use of additives (i.e., cyclodextrins) for enantiomeric separation as well as any covalent immobilization of the enzyme into the wall channels or on the electrode surface such as in the biosensor-based approaches. Hybrid polymer/graphene-based electrodes were end-channel coupled to the microfluidic system to improve the analytical performance. D-Met and D-Leu were successfully detected becoming this proof-of-the-concept a promising principle for the development of point-of-care (POC) devices for in situ screening of V. cholerae related diseases.
本文提出了一种电化学微流控策略，用于分离和对映体检测 D-甲硫氨酸（D-Met）和 D-亮氨酸（D-Leu）。这些 D-氨基酸（D-AAs）作为与霍乱弧菌引起的相关疾病有关的生物标志物。在单个布局微流控芯片（MC）上，与极低的生物样品消耗高度兼容，该策略允许对 D-氨基酸氧化酶（DAAO）和每种 D-AA 生物标志物之间的受控微流体 D-AA 分离以及 D-AA 生物标志物之间的特定反应，而无需使用添加剂（例如环糊精）进行对映体分离，也无需将酶共价固定在壁通道或电极表面上，如基于生物传感器的方法。混合聚合物/基于石墨烯的电极与微流控系统进行端通道耦合，以提高分析性能。成功检测到 D-Met 和 D-Leu，这一概念验证为开发用于现场筛查霍乱弧菌相关疾病的即时护理（POC）设备提供了有前景的原则。
