An Optical Biosensor for Continuous Glucose Monitoring in Animal Cell Cultures

Abstract

Biosensors for continuous glucose monitoring in bioreactors could provide a valuable tool for optimizing culture conditions in biotechnological applications. We have developed an optical biosensor for long-term continuous glucose monitoring and demonstrated a tight glucose level control during cell culture in disposable bioreactors. The in-line sensor is based on a commercially available oxygen sensor that is coated with cross-linked glucose oxidase (GOD). The dynamic range of the sensor was tuned by a hydrophilic perforated diffusion membrane with an optimized permeability for glucose and oxygen. The biosensor was thoroughly characterized by experimental data and numerical simulations, which enabled insights into the internal concentration profile of the deactivating by-product hydrogen peroxide. The simulations were carried out with a one-dimensional biosensor model and revealed that, in addition to the internal hydrogen peroxide concentration, the turnover rate of the enzyme GOD plays a crucial role for culture monitoring is an integral part of animal cell cultivation. For several culture parameters, in situ sensors exist; others are predominantly monitored off-line. One important cell culture parameter is glucose concentration. Despite many efforts, there is still a lack of in situ sensors for continuous glucose monitoring. Such biosensors could provide a valuable tool for optimizing culture conditions in biotechnological applications. In this contribution, the manufacture of a long-term stable optical glucose sensor is described which is used to demonstrate glucose level monitoring during cell culture in disposable bioreactors. The in situ sensor is based on a commercially available oxygen sensor that is coated with cross-linked glucose oxidase and a hydrophilic perforated diffusion membrane. Glucose was measured in shake flasks and wave bags with only minor drifts of the sensor sensitivity during batch and fed-batch fermentations.