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

Lasse Jannis Frey,Jan-Luca Lohse,Maximilian Breitfeld,Jan-Hendrik Grosch,Janina Bahnemann,Detlev Rasch,Rainer Krull,Gregor D Wehinger,David Vorländer,Hendrik Ostsieker

doi:10.1038/s41598-021-86654-9

Abstract

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.

Highlights

With the technological advances in 3D printing technology, which are associated with ever-increasing printing resolution, additive manufacturing is increasingly being used for rapid manufacturing of complex devices including microsystems development for laboratory applications
Edlich et al.[7] developed a 10 μL horizontal flow and passively gassed MBR system made of glass and PDMS with integrated online sensors for optical density (OD) and dissolved oxygen tension (DOT), which was used for the cultivation of Saccharomyces cerevisiae (S. cerevisiae)
This study presents for the first time a novel, modular design and reproducible setup of a 3D-printed micro bubble column reactor (3D-μBCR) with integrated sensor technology and a cultivation volume of 550 μL

Summary

Introduction

With the technological advances in 3D printing technology, which are associated with ever-increasing printing resolution, additive manufacturing is increasingly being used for rapid manufacturing of complex devices including microsystems development for laboratory applications. Various MBR systems have been developed for automated and parallel operation to enable, e.g., a realistic scale up/down of biotechnological processes[1], characterization of mammalian cell heterogeneity[2], and the screening of whole cell and biotransformation systems[3] This MBR technology allows researchers to obtain quantitative data concerning the most important process variables from a large number of simultaneously running cultivation approaches in real-time, with both high data density and a ccuracy[4,5,6]. This study presents for the first time a novel, modular design and reproducible setup of a 3D-printed micro bubble column reactor (3D-μBCR) with integrated sensor technology and a cultivation volume of 550 μL. Due to the small reaction volume, sampling and offline analysis related thereto is only possible to a limited extend

Methods

Results

Conclusion