Model-based analysis of biocatalytic processes and performance of microbioreactors with integrated optical sensors

Pichia pastoris has emerged in the past years as a promising host for recombinant protein and biopharmaceutical production. In the establishment of high cell density fed‐batch biomanufacturing, screening phase and early bioprocess development (based on microplates and shake flasks) still represent a bottleneck due to high‐cost and time‐consuming procedures as well as low experiment complexity. In the present work, a screening protocol developed for P. pastoris clone selection is implemented in a multiplexed microfluidic device with 15 μL cultivation chambers able to operate in perfusion mode and monitor dissolved oxygen content in the culture in a non‐invasive way. The setup allowed us to establish carbon‐limited conditions and evaluate strain responses to different input variables. Results from micro‐scale perfusion cultures are then compared with 1L fed‐batch fermentation. The best producer in terms of titer and productivity is rapidly identified after 12 h from inoculation and the results confirmed by lab‐scale fermentation. Moreover, the physiological analyses of the strains under different conditions suggested how more complex experimental conditions are achievable despite the relatively easy, straight‐forward, and cost‐effective experimental setup. Implementation and standardization of these micro‐scale protocols could reduce the demand for lab‐scale bioreactor cultivations thus accelerating the development of protein production processes.

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“…A contactless sensor spot was placed at the top of the chamber ensuring oxygen monitoring during cultivations. [ 16 ]…”

Section: Methodsmentioning

confidence: 99%

Microscale Perfusion‐Based Cultivation for Pichia pastoris Clone Screening Enables Accelerated and Optimized Recombinant Protein Production Processes

Totaro

Radoman

Schmelzer

et al. 2020

Biotechnology Journal

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“…The model predictions (identification of the reaction mechanism, kinetics and limiting factors) were independently confirmed for μLand mL-scale experiments. 103 Vobecká et al have recently presented an interesting work in which they present an enzymatic synthesis of cephalexin in a continuous-flow microfluidic device in aqueous two-phase system (ATPS) forming twophase slug flow in a microfluidic capillary as the reactionseparation environment. 104 ATPS consisted of 15 wt% of PEG 4000, 12 wt% of phosphates, and 73 wt% of water.…”

Section: Reaction Chemistry and Engineering Reviewmentioning

confidence: 99%

The rise of continuous flow biocatalysis – fundamentals, very recent developments and future perspectives

2020

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“…Therefore, a lot of computational models are focused on the optimization of fluid behavior in bioreactors, while a lot of biological models are focused on the cell culturing process in the bioreactors. Hence, considering the complexity of a bioreactor system, and the importance of controlling and predicting systems' behavior, different predictive computational models were proposed in the literature based on computational fluid dynamics (CFDs) [154][155][156][157][158][159][160][161].…”

Section: Mathematical Modeling and Computer Fluid Dynamics (Cfds)mentioning

confidence: 99%

“…The microbioreactors (µBRs) are relatively simple for modeling, due to the predictability and repeatability of fluid behavior, and initial parameters such as temperature, pH, amount of oxygen, and carbon dioxide in the system. There is a number of computational models for µBRs available in the literature, with a particular focus on the analyses and optimization of fluid behavior and mass transfer in the system, proposing efficient mixing methods for the µBR system [155], flow field and oxygen transport [156,157], mass transfer, fluid pressure and shear stress [158] and enzyme adsorption [159].…”

Section: Mathematical Modeling and Computer Fluid Dynamics (Cfds)mentioning

confidence: 99%

Cultivating Multidisciplinarity: Manufacturing and Sensing Challenges in Cultured Meat Production

Djisalov

Knežić

Podunavac

et al. 2021

Biology

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Meat cultivation via cellular agriculture holds great promise as a method for future food production. In theory, it is an ideal way of meat production, humane to the animals and sustainable for the environment, while keeping the same taste and nutritional values as traditional meat and having additional benefits such as controlled fat content and absence of antibiotics and hormones used in the traditional meat industry. However, in practice, there is still a number of challenges, such as those associated with the upscale of cultured meat (CM). CM food safety monitoring is a necessary factor when envisioning both the regulatory compliance and consumer acceptance. To achieve this, a multidisciplinary approach is necessary. This includes extensive development of the sensitive and specific analytical devices i.e., sensors to enable reliable food safety monitoring throughout the whole future food supply chain. In addition, advanced monitoring options can help in the further optimization of the meat cultivation which may reduce the currently still high costs of production. This review presents an overview of the sensor monitoring options for the most relevant parameters of importance for meat cultivation. Examples of the various types of sensors that can potentially be used in CM production are provided and the options for their integration into bioreactors, as well as suggestions on further improvements and more advanced integration approaches. In favor of the multidisciplinary approach, we also include an overview of the bioreactor types, scaffolding options as well as imaging techniques relevant for CM research. Furthermore, we briefly present the current status of the CM research and related regulation, societal aspects and challenges to its upscaling and commercialization.

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Model-based analysis of biocatalytic processes and performance of microbioreactors with integrated optical sensors

Cited by 14 publications

References 28 publications

Microscale Perfusion‐Based Cultivation for Pichia pastoris Clone Screening Enables Accelerated and Optimized Recombinant Protein Production Processes

Microscale Perfusion‐Based Cultivation for Pichia pastoris Clone Screening Enables Accelerated and Optimized Recombinant Protein Production Processes

The rise of continuous flow biocatalysis – fundamentals, very recent developments and future perspectives

Cultivating Multidisciplinarity: Manufacturing and Sensing Challenges in Cultured Meat Production

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