TY  - JOUR
A1  - Huck, Christina
A1  - Schiffels, Johannes
A1  - Herrera, Cony N.
A1  - Schelden, Maximilian
A1  - Selmer, Thorsten
A1  - Poghossian, Arshak
A1  - Baumann, Marcus
A1  - Wagner, Patrick
A1  - Schöning, Michael Josef
T1  - Metabolic responses of Escherichia coli upon glucose pulses captured by a capacitive field-effect sensor
JF  - Physica Status Solidi (A)
N2  - Living cells are complex biological systems transforming metabolites taken up from the surrounding medium. Monitoring the responses of such cells to certain substrate concentrations is a challenging task and offers possibilities to gain insight into the vitality of a community influenced by the growth environment. Cell-based sensors represent a promising platform for monitoring the metabolic activity and thus, the “welfare” of relevant organisms. In the present study, metabolic responses of the model bacterium Escherichia coli in suspension, layered onto a capacitive field-effect structure, were examined to pulses of glucose in the concentration range between 0.05 and 2 mM. It was found that acidification of the surrounding medium takes place immediately after glucose addition and follows Michaelis–Menten kinetic behavior as a function of the glucose concentration. In future, the presented setup can, therefore, be used to study substrate specificities on the enzymatic level and may as well be used to perform investigations of more complex metabolic responses. Conclusions and perspectives highlighting this system are discussed.
Y1  - 2013
U6  - http://dx.doi.org/10.1002/pssa.201200900
SN  - 0031-8965
VL  - 210
IS  - 5
SP  - 926
EP  - 931
PB  - Wiley-VCH
CY  - Weinheim
ER  - 
TY  - JOUR
A1  - Bäcker, Matthias
A1  - Rakowski, D.
A1  - Poghossian, Arshak
A1  - Biselli, Manfred
A1  - Wagner, Patrick
A1  - Schöning, Michael Josef
T1  - Chip-based amperometric enzyme sensor system for monitoring of bioprocesses by flow-injection analysis
JF  - Journal of Biotechnology
N2  - 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.
Y1  - 2013
U6  - http://dx.doi.org/10.1016/j.jbiotec.2012.03.014
SN  - 0168-1656
VL  - 163
IS  - 4
SP  - 371
EP  - 376
PB  - Elsevier
CY  - Amsterdam
ER  -