Bioprocess characterization at the micro-scale: Optical sensor integration in a novel capillary-wave micro-bioreactor

Due to the high demand of new biopharmaceuticals and bioproducts, the development of new cultivation platforms for high-throughput screenings, cell-based assays and bioprocess development is of high interest. Therefore, micro-bioreactors (MBRs) are a promising alternative to conventional cultivation platforms like shake flasks due to their minimal volume, sensor integration and high ability for automatization and parallelization. Especially, MBRs with a volume below 10 µL can reduce the amount of needed testing substances for cell-based assays, which is advantageous mostly for testing new biopharmaceuticals with limited availability. However, characterization of a cell culture in the lower micro-liter scale is challenging due to the limited space and the insufficient volume for sampling and offline analysis. Optical sensors are one suitable possibility to close this gap. Therefore, a novel capillary-wave micro-bioreactor (cwMBR) with a working volume of 7 µL and optical sensors for biomass, glucose, oxygen, pH and fluorescence intensity measurement was developed. Please click Additional Files below to see the full abstract

3D-printed micro bubble column reactor with integrated microsensors for biotechnological applications: From design to evaluation

With the technological advances in 3D printing technology, which are associated with ever-increasing printing resolution, additive manufacturing is now increasingly being used for rapid manufacturing of complex devices including microsystems development for laboratory applications. Personalized experimental devices or entire bioreactors of high complexity can be manufactured within few hours from start to finish. This study presents a customized 3D-printed micro bubble column reactor (3D-µBCR), which can be used for the cultivation of microorganisms (e.g., Saccharomyces cerevisiae) and allows online-monitoring of process parameters through integrated microsensor technology. The modular 3D-µBCR achieves rapid homogenization in less than 1 s and high oxygen transfer with kLa values up to 788 h−1 and is able to monitor biomass, pH, and DOT in the fluid phase, as well as CO2 and O2 in the gas phase. By extensive comparison of different reactor designs, the influence of the geometry on the resulting hydrodynamics was investigated. In order to quantify local flow patterns in the fluid, a three-dimensional and transient multiphase Computational Fluid Dynamics model was successfully developed and applied. The presented 3D-µBCR shows enormous potential for experimental parallelization and enables a high level of flexibility in reactor design, which can support versatile process development. © 2021, The Author(s)

3D-printed micro bubble column reactor with integrated microsensors for biotechnological applications: from design to evaluation

With the technological advances in 3D printing technology, which are associated with ever-increasing printing resolution, additive manufacturing is now increasingly being used for rapid manufacturing of complex devices including microsystems development for laboratory applications. Personalized experimental devices or entire bioreactors of high complexity can be manufactured within few hours from start to finish. This study presents a customized 3D-printed micro bubble column reactor (3D-µBCR), which can be used for the cultivation of microorganisms (e.g., Saccharomyces cerevisiae) and allows online-monitoring of process parameters through integrated microsensor technology. The modular 3D-µBCR achieves rapid homogenization in less than 1 s and high oxygen transfer with kLa values up to 788 h-1 and is able to monitor biomass, pH, and DOT in the fluid phase, as well as CO2 and O2 in the gas phase. By extensive comparison of different reactor designs, the influence of the geometry on the resulting hydrodynamics was investigated. In order to quantify local flow patterns in the fluid, a three-dimensional and transient multiphase Computational Fluid Dynamics model was successfully developed and applied. The presented 3D-µBCR shows enormous potential for experimental parallelization and enables a high level of flexibility in reactor design, which can support versatile process development

Microsensor in Microbioreactors: Full Bioprocess Characterization in a Novel Capillary-Wave Microbioreactor

Microbioreactors (MBRs) with a volume below 1 mL are promising alternatives to established cultivation platforms such as shake flasks, lab-scale bioreactors and microtiter plates. Their main advantages are simple automatization and parallelization and the saving of expensive media components and test substances. These advantages are particularly pronounced in small-scale MBRs with a volume below 10 µL. However, most described small-scale MBRs are lacking in process information from integrated sensors due to limited space and sensor technology. Therefore, a novel capillary-wave microbioreactor (cwMBR) with a volume of only 7 µL has the potential to close this gap, as it combines a small volume with integrated sensors for biomass, pH, dissolved oxygen (DO) and glucose concentration. In the cwMBR, pH and DO are measured by established luminescent optical sensors on the bottom of the cwMBR. The novel glucose sensor is based on a modified oxygen sensor, which measures the oxygen uptake of glucose oxidase (GOx) in the presence of glucose up to a concentration of 15 mM. Furthermore, absorbance measurement allows biomass determination. The optical sensors enabled the characterization of an Escherichia coli batch cultivation over 8 h in the cwMBR as proof of concept for further bioprocesses. Hence, the cwMBR with integrated optical sensors has the potential for a wide range of microscale bioprocesses, including cell-based assays, screening applications and process development

A fully online sensor-equipped, disposable multiphase microbioreactor as a screening platform for biotechnological applications

Abstract A new disposable, multiphase, microbioreactor (MBR; with a working volume of 550 μl) equipped with online sensors is presented for biotechnological screening research purposes owing to its high‐throughput potential. Its design and fabrication, online sensor integration, and operation are described. During aerobic cultivation, sufficient oxygen supply is the most important factor that influences growth and product formation. The MBR is a microbubble column bioreactor (μBC), and the oxygen supply was realized by active pneumatic bubble aeration, ensuring sufficient volumetric liquid‐phase mass transfer (kLa) and proper homogenization of the cultivation broth. The μBC was equipped with miniaturized sensors for the pH, dissolved oxygen, optical density and glucose concentration that allowed real‐time online monitoring of these process variables during cultivation. The challenge addressed here was the integration of sensors in the limited available space. The MBR was shown to be a suitable screening platform for the cultivation of biological systems. Batch cultivations of Saccharomyces cerevisiae were performed to observe the variation in the process variables over time and to show the robustness and operability of all the online sensors in the MBR