Rentschler Biopharma SE, Laupheim, Germany.
Department of Biotechnology, Mannheim University of Applied Sciences, Mannheim, Germany.
Methods Mol Biol. 2020;2095:319-333. doi: 10.1007/978-1-0716-0191-4_19.
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.
用于生物反应器中连续葡萄糖监测的生物传感器可为优化生物技术应用中的培养条件提供有价值的工具。我们开发了一种用于长期连续葡萄糖监测的光学生物传感器，并在一次性生物反应器中的细胞培养过程中证明了严格的葡萄糖水平控制。在线传感器基于市售的氧传感器，该氧传感器涂有交联的葡萄糖氧化酶（GOD）。传感器的动态范围通过具有优化的葡萄糖和氧气渗透性的亲水穿孔扩散膜进行调节。通过实验数据和数值模拟对生物传感器进行了全面表征，这使我们深入了解了失活副产物过氧化氢的内部浓度分布。模拟是使用一维生物传感器模型进行的，结果表明，除了内部过氧化氢浓度外，酶 GOD 的周转率对于培养监测也起着至关重要的作用。在线传感器是动物细胞培养的一个组成部分。对于许多培养参数，都存在原位传感器；而其他参数主要是离线监测。一个重要的细胞培养参数是葡萄糖浓度。尽管做了很多努力，但仍缺乏用于连续葡萄糖监测的原位传感器。这种生物传感器可为优化生物技术应用中的培养条件提供有价值的工具。在本贡献中，描述了一种长期稳定的光学葡萄糖传感器的制造方法，该传感器用于在一次性生物反应器中的细胞培养过程中证明葡萄糖水平监测。该原位传感器基于市售的氧传感器，该氧传感器涂有交联的葡萄糖氧化酶和亲水穿孔扩散膜。在分批和补料分批发酵过程中，仅传感器灵敏度略有漂移，即可在摇瓶和波袋中测量葡萄糖。
