Characterization of hydrodynamics and volumetric power input in microtiter plates for the scale‐up of downstream operations

Montes‐Serrano, Ignacio; Satzer, Peter; Jungbauer, Alois; Dürauer, Astrid

doi:10.1002/bit.27983

Cited by 8 publications

(3 citation statements)

References 37 publications

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“…However, Montes‐Serano et al. showed that for increasing filling volumes in 6‐, 24‐, and 96‐well plates a decrease in volumetric power input due to the change of surface to volume ratio can be noted [ 32 ]. The same effect holds true for 48‐well plates.…”

Section: Resultsmentioning

confidence: 99%

Insights into Streptomyces coelicolor A3(2) growth and pigment formation with high‐throughput online monitoring

Finger

Sentek

Hartmann

et al. 2022

Engineering in Life Sciences

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Streptomyces species are intensively studied for their ability to produce a variety of natural products. However, conditions influencing and leading to product formation are often not completely recognized. Therefore, in this study, high-throughput online monitoring is presented as a powerful tool to gain indepth understanding of the cultivation of the model organism Streptomyces coelicolor A3(2). Through online measurements of oxygen transfer rate and autofluorescence, valuable information about availability of nutrients and product formation patterns of the pigments actinorhodin and undecylprodigiosin can be obtained and explained. Therefore, it is possible to determine the onset of pigmentation and to study in detail the influencing factors thereof. One factor identified in this study is the filling volume of the cultivation vessel. Slight variations led to varying pigmentation levels. By combining optical and metabolic online monitoring techniques, the correlation of the filling volume with pigmentation could be explained as a result of different growth trajectories caused by varying specific power inputs and their influence on the pellet formation of the filamentous system. Finally, experiments with the addition of supernatant from unpigmented and pigmented cultures could highlight the applicability of the presented approach to study quorum sensing and cell-cell interaction.

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Section: Resultsmentioning

confidence: 99%

Insights into Streptomyces coelicolor A3(2) growth and pigment formation with high‐throughput online monitoring

Finger

Sentek

Hartmann

et al. 2022

Engineering in Life Sciences

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show abstract

“…However, this approach becomes difficult for larger scale differences, because certain factors, such as the power input per volume ratio (P/V), are impractical to be kept constant. ( Marques et al, 2010 ; Tajsoleiman et al, 2019 ; Montes‐Serrano et al, 2021 ). For ADCs, no systematic approach has been reported and scale-comparability is assessed based on constant ADC quality attributes, such as the DAR or aggregate level ( Hutchinson et al, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Kinetic studies and CFD-based reaction modeling for insights into the scalability of ADC conjugation reactions

Weggen

Seidel

Bean³

et al. 2023

Front. Bioeng. Biotechnol.

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The manufacturing of antibody-drug conjugates (ADCs) involves the addition of a cytotoxic small-molecule linker-drug (= payload) to a solution of functionalized antibodies. For the development of robust conjugation processes, initially small-scale reaction tubes are used which requires a lot of manual handling. Scale-up to larger reaction vessels is often knowledge-driven and scale-comparability is solely assessed based on final product quality which does not account for the dynamics of the reaction. In addition, information about the influence of process parameters, such as stirrer speed, temperature, or payload addition rates, is limited due to high material costs. Given these limitations, there is a need for a modeling-based approach to investigate conjugation scale-up. In this work, both experimental kinetic studies and computational fluid dynamics (CFD) conjugation simulations were performed to understand the influence of scale and mixing parameters. In the experimental part, conjugation kinetics in small-scale reaction tubes with different mixing types were investigated for two ADC systems and compared to larger bench-scale reactions. It was demonstrated that more robust kinetics can be achieved through internal stirrer mixing instead of external mixing devices, such as orbital shakers. In the simulation part, 3D-reactor models were created by coupling CFD-models for three large-scale reaction vessels with a kinetic model for a site-specific conjugation reaction. This enabled to study the kinetics in different vessels, as well as the effect of process parameter variations in silico. Overall, it was found that for this conjugation type sufficient mixing can be achieved at all scales and the studied parameters cause only deviations during the payload addition period. An additional time-scale analysis demonstrated to aid the assessment of mixing effects during ADC process scale-up when mixing times and kinetic rates are known. In summary, this work highlights the benefit of kinetic models for enhanced conjugation process understanding without the need for large-scale experiments.

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“…To calculate the volumetric power input from CFD simulations, the relationship to the energy dissipation rate was used: where P, V L and ε are the power input in W, the liquid volume in m 3 and the energy dissipation rate in W/kg. This approach has been used before for the simulation of shaken system in general 15 – 17 and shake flasks specifically 19 . The total energy dissipation consists of the dissipation of the mean flow and dissipation caused by the turbulence.…”

Section: Methodsmentioning

confidence: 99%

Validation of computational fluid dynamics of shake flask experiments at moderate viscosity by liquid distributions and volumetric power inputs

Dinter,

Gumprecht,

Menze

et al. 2024

Sci Rep

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Computational fluid dynamics (CFD) has recently become a pivotal tool in the design and scale-up of bioprocesses. While CFD has been extensively utilized for stirred tank reactors (STRs), there exists a relatively limited body of literature focusing on CFD applications for shake flasks, almost exclusively concentrated on fluids at waterlike viscosity. The importance of CFD model validation cannot be overstated. While techniques to elucidate the internal flow field are necessary for model validation in STRs, the liquid distribution, caused by the orbital shaking motion of shake flasks, can be exploited for model validation. An OpenFOAM CFD model for shake flasks has been established. Calculated liquid distributions were compared to suitable, previously published experimental data. Across a broad range of shaking conditions, at waterlike and moderate viscosity (16.7 mPa∙s), the CFD model's liquid distributions align excellently with the experimental data, in terms of overall shape and position of the liquid relative to the direction of the centrifugal force. Additionally, the CFD model was used to calculate the volumetric power input, based on the energy dissipation. Depending on the shaking conditions, the computed volumetric power inputs range from 0.1 to 7 kW/m3 and differed on average by 0.01 kW/m3 from measured literature data.

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Characterization of hydrodynamics and volumetric power input in microtiter plates for the scale‐up of downstream operations

Cited by 8 publications

References 37 publications

Insights into Streptomyces coelicolor A3(2) growth and pigment formation with high‐throughput online monitoring

Insights into Streptomyces coelicolor A3(2) growth and pigment formation with high‐throughput online monitoring

Kinetic studies and CFD-based reaction modeling for insights into the scalability of ADC conjugation reactions

Validation of computational fluid dynamics of shake flask experiments at moderate viscosity by liquid distributions and volumetric power inputs

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