Institute of Nano- and Biotechnologies-INB, Aachen University of Applied Sciences, Campus Jülich, Germany.
J Biotechnol. 2013 Feb 20;163(4):371-6. doi: 10.1016/j.jbiotec.2012.03.014. Epub 2012 Mar 23.
A microfluidic chip integrating amperometric enzyme sensors for the detection of glucose, glutamate and glutamine in cell-culture fermentation processes has been developed. The enzymes glucose oxidase, glutamate oxidase and glutaminase were immobilized by means of cross-linking with glutaraldehyde on platinum thin-film electrodes integrated within a microfluidic channel. The biosensor chip was coupled to a flow-injection analysis system for electrochemical characterization of the sensors. The sensors have been characterized in terms of sensitivity, linear working range and detection limit. The sensitivity evaluated from the respective peak areas was 1.47, 3.68 and 0.28 μAs/mM for the glucose, glutamate and glutamine sensor, respectively. The calibration curves were linear up to a concentration of 20 mM glucose and glutamine and up to 10 mM for glutamate. The lower detection limit amounted to be 0.05 mM for the glucose and glutamate sensor, respectively, and 0.1 mM for the glutamine sensor. Experiments in cell-culture medium have demonstrated a good correlation between the glutamate, glutamine and glucose concentrations measured with the chip-based biosensors in a differential-mode and the commercially available instrumentation. The obtained results demonstrate the feasibility of the realized microfluidic biosensor chip for monitoring of bioprocesses.
已开发出一种微流控芯片，该芯片集成了安培酶传感器，可用于检测细胞培养发酵过程中的葡萄糖、谷氨酸和谷氨酰胺。通过戊二醛交联将葡萄糖氧化酶、谷氨酸氧化酶和谷氨酰胺酶固定在集成在微流道内的铂薄膜电极上。将生物传感器芯片与流动注射分析系统耦合，以对传感器进行电化学表征。根据各自的峰面积评估了传感器的灵敏度、线性工作范围和检测限。葡萄糖、谷氨酸和谷氨酰胺传感器的灵敏度分别为 1.47、3.68 和 0.28 μAs/mM。校准曲线在葡萄糖和谷氨酰胺浓度高达 20 mM 以及谷氨酸浓度高达 10 mM 时呈线性。葡萄糖和谷氨酸传感器的检测下限分别为 0.05 mM，谷氨酰胺传感器的检测下限为 0.1 mM。在细胞培养液中的实验表明，基于芯片的生物传感器在差分模式下测量的谷氨酸、谷氨酰胺和葡萄糖浓度与市售仪器之间具有良好的相关性。所获得的结果证明了所实现的微流控生物传感器芯片用于监测生物过程的可行性。
