Al-Halhouli Ala'aldeen, Demming Stefanie, Alahmad Laila, LIobera Andreu, Büttgenbach Stephanus
Institute of Microtechnology, Technische Universität Braunschweig, Langer Kamp 8, 38106 Braunschweig, Germany.
CentreNacional de Microelectrònica (IMB-CNM, CSIC), 08193 Barcelona, Spain.
Sensors (Basel). 2014 Aug 25;14(9):15749-59. doi: 10.3390/s140915749.
This paper presents two PDMS photonic biosensor designs that can be used for continuous monitoring of glucose concentrations. The first design, the internally immobilized sensor, consists of a reactor chamber, micro-lenses and self-alignment structures for fiber optics positioning. This sensor design allows optical detection of glucose concentrations under continuous glucose flow conditions of 33 µL/h based on internal co-immobilization of glucose oxidase (GOX) and horseradish peroxidase (HRP) on the internal PDMS surface of the reactor chamber. For this design, two co-immobilization methods, the simple adsorption and the covalent binding (PEG) methods were tested. Experiments showed successful results when using the covalent binding (PEG) method, where glucose concentrations up to 5 mM with a coefficient of determination (R2) of 0.99 and a limit of detection of 0.26 mM are detectable. The second design is a modified version of the internally immobilized sensor, where a microbead chamber and a beads filling channel are integrated into the sensor. This modification enabled external co-immobilization of enzymes covalently onto functionalized silica microbeads and allows binding a huge amount of HRP and GOX enzymes on the microbeads surfaces which increases the interaction area between immobilized enzymes and the analyte. This has a positive effect on the amount and rate of chemical reactions taking place inside the chamber. The sensor was tested under continuous glucose flow conditions and was found to be able to detect glucose concentrations up to 10 mM with R2 of 0.98 and a limit of detection of 0.7 mM. Such results are very promising for the application in photonic LOC systems used for online analysis.
本文介绍了两种可用于连续监测葡萄糖浓度的聚二甲基硅氧烷(PDMS)光子生物传感器设计。第一种设计是内部固定化传感器,由一个反应腔、微透镜和用于光纤定位的自对准结构组成。这种传感器设计允许在33 μL/h的连续葡萄糖流动条件下,基于葡萄糖氧化酶(GOX)和辣根过氧化物酶(HRP)在反应腔内部PDMS表面的共固定化,对葡萄糖浓度进行光学检测。对于这种设计,测试了两种共固定化方法,即简单吸附法和共价结合(PEG)法。实验表明,使用共价结合(PEG)法时取得了成功结果,可检测到高达5 mM的葡萄糖浓度,测定系数(R2)为0.99,检测限为0.26 mM。第二种设计是内部固定化传感器的改进版本,其中将一个微珠腔和一个珠子填充通道集成到传感器中。这种改进使得能够将酶共价地外部共固定到功能化的二氧化硅微珠上,并允许在微珠表面结合大量的HRP和GOX酶,这增加了固定化酶与分析物之间的相互作用面积。这对反应腔内发生的化学反应的量和速率有积极影响。该传感器在连续葡萄糖流动条件下进行了测试,发现能够检测到高达10 mM的葡萄糖浓度,R2为0.98,检测限为0.7 mM。这些结果对于用于在线分析的光子微流控系统的应用非常有前景。