CFD simulations are widely used to quantify the mixing performance of stirred tanks for various applications in chemical engineering and biotechnology. Due to advances in GPU computing, these simulations increasingly employ Large Eddy Simulation (LES), which explicitly resolves the dynamics of large-scale turbulence. Although such simulations are fully deterministic and therefore theoretically reproducible, small numerical variations induced by round-off errors, floating-point arithmetic, and differences in the distribution and ordering of operations in parallel computing lead to separation of trajectories i.e., different flow-field evolutions and consequently to significant run-to-run variability in predicted mixing times, even on the same hardware architecture. This work investigates the impact of repeated simulations, in the form of a case study, on the mixing-time distribution observed in a 30L stirred tank reactor using two commercial CFD packages operating with representative, production-level solver configurations. The analysis does not aim to assess the general performance of numerical method classes, but rather to quantify run-to-run variability under fixed solver settings and to compare the resulting numerical distributions to experimental variability. The results demonstrate that numerical variability is of comparable magnitude to the experimental spread, highlighting the necessity to treat LES-derived metrics as statistical ensembles rather than deterministic values. It is concluded that the reporting of confidence intervals is essential for methodological rigour in LES-based mixing studies.

Background - An effective treatment of symptomatic Atrial fibrillation (AF) is the second-generation cryoballoon (CB2) based pulmonary vein isolation (PVI). However, this technique is associated with an increased exposure to contrast media and little is known about postprocedural acute kidney injury (AKI). Objective - We evaluated the incidence, characteristics and risk factors of contrast-associated AKI after CB2-based catheter ablation in a large patient cohort. Methods - 444 patients with symptomatic drug-refractory paroxysmal AF (PAF, n=236 [53%]) or persistent AF (n=208) who underwent CB2-based PVI were included. The study group was divided into four groups based on the estimated glomerular filtration rate (eGFR) documented the day before the procedure. Serum creatinine levels were measured before and 48 hours after the procedure. AKI was assessed using the KDIGO guidelines. Results - The overall incidence of AKI was 2.9% (13/444). Among the AKI group, the majority were in CKD stage 4 (2/7, 28.6%), followed by CKD stage 3 (7/111; 6.3%), CKD stage 1(1/70; 1.4%) and CKD stage 2 (3/256; 1.2%). Baseline CKD stage (HR 1.3659, 95% CI 0.05107 - 36.53, p = 0.033) was identified as independent predictor of AKI. Arterial hypertension was prevalent in the AKI group. Conclusions - The overall incidence of postcontrast AKI in CB2-based ablation of AF was low. Cryoablation PVI is a safe therapeutic option for individuals with CKD stages 1 and 2. However, in patients with CKD stages 3 and 4, it is associated with an increased incidence of AKI.

Utilizing pressure as a process parameter can make biotechnological processes more efficient and attractive compared to established ones. This paper presents a high-pressure reactor setup for enzymatically catalyzed gas-liquid reactions, which can be operated up to 15.0 MPa. The reactor is equipped with optical measurement technology for inline and in situ monitoring of the oxygen concentration under high-pressure conditions. The setup is characterized by assessing the influence of the process parameter pressure on the conversion of the glucose oxidation to d-glucono-delta-lactone by immobilized glucose oxidase. The study demonstrates that the increased oxygen availability due to higher solubility reduces the reaction time in a batch reactor from 270 to 90 min.

A new model to predict the breakup of gaseous bubbles in a continuous liquid phase is developed. In the model each bubble is modelled as a spring-damper system, namely a Kelvin-Voigt element, while the outer force is derived by a Lagrangian analysis determining the largest stretching rate of the flow field below. The developed model is based on physical principles and no further arbitrary parameters have to be adjusted. Each bubble is observed on its way through the bubbly flow individually, taking into account its history along its respective path. With the implemented model numerical simulations in a wide range of scales are conducted, ranging from the laboratory scale of a vessel of 3 L to the large industrial scale of 15 m(3). The simplicity of the model allows for a good cost to benefit ratio. In the present work, the achieved results are compared to experimental data obtained from optical measurements in a replica of a 200 L aerated stirred tank reactor for various stirrer frequencies.

The Villermaux-Dushman protocol with UV-Vis analytics is an established tool to characterize the global micro mixing performance in process equipment. The local mixing process is described by a micro mixing model (incorporation model), originally designed for turbulent flows. The novel imaging UV-Vis spectroscopy used in this work uncovers locally resolved micro mixing phenomena in a laminar split-and-recombine (SAR) mixer unit manufactured from selective laser-induced etching (SLE). The local absorbance is recorded with a high spatial resolution camera through a telecentric lens and post-processed into local concentration fields of components. The method unveils discrepancies of the micro mixing time determined from the conventional incorporation model in laminar flow and the locally recorded mixing process. The micro mixing time rather needs to be seen as a mean micro mixing time instead. Furthermore, the spatial information obtained by the imaging UV-Vis spectroscopy gives insight into local micro mixing and selectivity, yielding new approaches to equipment optimization.

During the scale-up of biopharmaceutical production processes, insufficiently predictable performance losses may occur alongside gradients and heterogeneities. To overcome such performance losses, tools are required to explain, predict, and ultimately prohibit inconsistencies between laboratory and commercial scale. In this work, we performed CHO fed-batch cultivations in the single multicompartment bioreactor (SMCB), a new scale-down reactor system that offers new access to study large-scale heterogeneities in mammalian cell cultures. At volumetric power inputs of 20.4-1.5 W m-3, large-scale characteristics like long mixing times and dissolved oxygen (DO) heterogeneities were mimicked in the SMCB. Compared to a reference bioreactor (REFB) set-up, the conditions in the SMCB provoked an increase in lactate accumulation of up to 87%, an increased glucose uptake, and reduced viable cell concentrations in the stationary phase. All are characteristic for large-scale performance. The unique possibility to distinguish between the effects of changing power inputs and observed heterogeneities provided new insights into the potential reasons for altered product quality attributes. Apparently, the degree of galactosylation in the evaluated glycan patterns changed primarily due to the different power inputs rather than the provoked heterogeneities. The SMCB system could serve as a potent tool to provide new insights into scale-up behavior and to predict cell line-specific drawbacks at an early stage of process development.

Aerated stirred tank reactors are widely used in bio-process engineering and pharmaceutical industries. To supply the organisms with oxygen and control the pH value, oxygen is transferred from air bubbles into the liquid phase, and, at the same time, carbon dioxide is stripped from the liquid phase with the same gas bubbles. The volumetric mass transfer coefficients for oxygen and carbon dioxide are, therefore, of crucial importance for the design and scale-up of aerated stirred tank reactors. In this experimental work, the volumetric mass transfer coefficients for oxygen and carbon dioxide are investigated simultaneously to study their mutual influence. The mass transfer performance for oxygen and carbon dioxide is conducted in stirred tank reactors on the 3 L laboratory scale, 30 L pilot scale, and 15,000 L production scale. First, the influence of dissolved carbon dioxide on the oxygen mass transfer performance is investigated in a 30 L pilot scale stirred tank reactor. The results show that the volumetric mass transfer coefficient of oxygen is not affected by the concentration of dissolved carbon dioxide, but the total mass flux of oxygen decreases with increasing carbon dioxide concentration due to the decreasing partial pressure difference. With rising gassing rate and volumetric power input, both mass transfer coefficients for oxygen and carbon dioxide show the same increasing trend. Although this trend can also be observed when scaling down to the 3 L laboratory scale reactor, a significantly different effect must be considered for the scale-up to the 15,000 L industrial scale reactor. The limited absorption capacity for carbon dioxide of the gas bubbles during the long residence time in the industrial scale reactor is noticeable here, which is why the specific interfacial area is of negligible importance. This effect is used to develop a method for independent control of oxygen and carbon dioxide mass transfer performance on an industrial scale and to increase the mass transfer performance for carbon dioxide by up to 25%.

This study employs Lagrangian Sensor Particles (LSPs) with a diameter of 40 mm equipped with a pressure sensor to investigate cell lifelines in a 15,000L stirred tank reactor (STR) with three Elephant Ear impellers. The Stokes number of the LSPs is approx. 0.004 on a macro-scale. The vertical probability of presence, axial velocity profiles, circulation time distributions, and residence time distributions are quantified to analyze single-phase mixing heterogeneities, detect hydrodynamic compartments and conduct a Lagrangian regime analysis. Results reveal a similarly distributed probability of presence in the vertical reactor center but emphasize the LSP’s sensitivity to fluctuating densities. Axial velocity distributions illustrate characteristic impeller-induced flow patterns, and circulation time distributions identify three compartments with comparatively shorter times in the axial center. Residence time distributions exhibit a similar compartmentalized profile. Moreover, the study estimates a potential oxygen deprivation zone for CHO cells in the upper compartment and demonstrates the LSP’s efficacy in characterizing impeller systems. Contrary to literature, the ratio of examined global mixing times to circulation times is 1.0, highlighting macro-scale mixing. The research underscores that LSPs offer crucial insights into industrial-scale STRs, specifically for determining hydrodynamic compartments without having optical access.

The reliable transfer of bioprocesses from single-use bioreactors (SUBs) of different scales to conventional stainless steel stirred-tank bioreactors is of steadily growing interest. In this publication, a scale-up study for SUBs with volumes of 200 L and 2000 L and the transfer to an industrial-scale conventional stainless steel stirred-tank bioreactor with a volume of 15,000 L is presented. The scale-up and transfer are based on a comparison of mixing times and the modeling of volumetric mass transfer coefficients kLa, measured in all three reactors in aqueous PBS/Kolliphor solution. The mass transfer coefficients are compared with the widely used correlation of van’t Riet at constant stirrer tip speeds. It can be shown that a van’t Riet correlation enables a robust and reliable prediction of mass transfer coefficients on each scale for a wide range of stirrer tip speeds and aeration rates. The process transfer from single-use bioreactors to conventional stainless steel stirred-tank bioreactors is proven to be uncritical concerning mass transfer performance. This provides higher flexibility with respect to bioreactor equipment considered for specific processes.

The utilization of oxidases suffers from insufficient oxygen availability due to its low solubility in gas-liquid multiphase systems. To overcome this limitation, oxygen is continuously supplied to the reaction medium during the course of the reaction, however, it causes significant gas consumption. In this study, a novel experimental set-up was established by installing a sintered frit porous sparger in the SpinChem® rotating bed reactor. Covalently immobilized glucose oxidase on epoxy-functionalized carriers was used in the rotating bed reactor with an activity yield of the immobilization of over 98%. For fine bubble aeration, the volumetric mass transfer coefficient kLa (67.3h-1) tripled compared to macrobubble aeration (22.5h-1), while the volume-specific aeration rate remained constant. A maximum reaction yield of 96.5% was reached. The reaction rate was improved by a factor of 12.9 for the fine bubble aeration compared to macrobubble aeration under identical conditions. With fine bubble aeration, the oxygen consumption was reduced by 87.5% over macrobubble aeration, establishing the same enzyme-specific reaction rates at a comparable kLa. Validation was carried out in repetitive batches, achieving the same glucose oxidase specific activity. This study demonstrates the advantages of fine bubble aeration and presents promising results for addressing the sustainable biotransformation processes.

The growing demand for bio-pharmaceuticals necessitates improved methods for the characterization of stirred tank reactors (STRs) and their mixing heterogeneities. Traditional Eulerian measurement approaches fall short, culminating in the use of Lagrangian Sensor Particles (LSPs) to map large-scale STRs and track the lifelines of microorganisms such as Chinese Hamster Ovary cells. This study investigates the hydrodynamic characteristics of LSPs, specifically examining the effects that the size and position of the Center of Mass (CoM) have on their flow-following capabilities. Two Lagrangian Particle (LP) designs are evaluated, one with the CoM and a Geometric Center aligned, and another with a shifted CoM. The experimental study is conducted in a rectangular vessel filled with deionized water featuring a stationary circular flow. Off-center LPs exhibit higher velocities, an increased number of floor contacts, and moreover, a less homogeneous particle probability of presence within the vessel compared to LPs with CoM and Geometric Center aligned. Lattice Boltzmann Large Eddy Simulations provide complementary undisturbed fluid velocity data for the calculation of the Stokes number St. Building upon these findings, differences in the Stokes number St between the two LP variants of ΔSt = 0.01 (25 mm LP) and ΔSt = 0.13 (40 mm LP) are calculated, highlighting the difference in flow behavior. Furthermore, this study offers a more representative calculation of particle response time approach, as the traditional Stokes number definition does not account for non-homogeneous particles, resulting in an alternative Stokes number (ΔStalt = 0.84 (25 mm LP) and ΔStalt = 2.72 (40 mm LP)). This study contributes to the improved characterization of STRs through the use of Lagrangian Sensor Particles. Results highlight the implications the internal mass distribution has on LSP design, offering crucial considerations for researchers in the field.

Mammalian cell cultivation in pharmaceutical industry can last up to units of weeks and requires proper transport of nutrients and oxygen for cell growth and production. Given the long time period, cells experience flow fields from all bioreactor’s zones, where the energy dissipation rate (ε) varies substantially. Shear sensitive micro-probes with size comparable to cells and Kolmogorov eddies are used for the determination of the maximum hydrodynamic stress (τmax) in bioreactors. For the very first time, the micro-probe method is applied successfully not only to laboratory (3L) and pilot scale (80L and 200L), but also to industrial production scale bioreactor (12,500L) with Rushton turbine and pitched-blade (RT-PB) impeller configuration. Experimentally obtained data are used for the validation of comprehensive CFD scale-up study, using the Lattice-Boltzmann large eddy simulation (LB-LES) method. Besides τmax, this work also focuses on the study of mixing time and flow field attributes.

Zielsetzung Evaluation der Kombination von 4D Fluss MRT und Computational Fluid Dynamics (CFD) zur nicht-invasiven Abschätzung des portosystemischen Druckgradienten (PSG) bei Patienten mit transjugulärem intrahepatischen portosystemischen Shunt (TIPS).

The detection of compartments and dead zones as well as the estimation of the mixing efficiency in stirred tanks are of vital interest for a variety of biochemical and chemical processes. Here, numerically derived time-dependent 3D fluid velocity fields of a stirred tank reactor are computed using the Lattice Boltzmann Method. Mixing in the stirred tank reactor is analysed by means of Lagrangian Coherent Structures which allow to unravel the mixing states of complex flows. This Lagrangian analysis is achieved by computing Finite Time Lyapunov Exponents and applying recent trajectory-based network methods on the three-dimensional flow. The results reveal a zone of low interaction in the upper region of the stirred tank reactor and five additional transient compartments. The trajectory-based network analysis detects low cross-mixing of fluid parcels between different compartments but a very high mixing of fluid parcels inside of each compartment. This high interaction is also found in an analysis of the Finite Time Lyapunov Mixing Intensity. Time-averaging of the fluid velocity field prior to the Lagrangian analysis is considered to extract the most influential Lagrangian Coherent Structures.

One of the key elements in stirred tank reactor set-ups is the submersed aeration system. Aeration with microbubbles provides high gas utility coupled with prolonged enzyme stability by lowering the interfacial area renewing rate depending on the aeration mode.