Continuous Bioprocess Development: Methods for Control and Characterization of the Biological System

Neubauer, Peter; Bournazou, Mariano Nicolás Cruz

doi:10.1002/9783527699902.ch1

Cited by 3 publications

(3 citation statements)

References 111 publications

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“…In this review, we summarize how continuous cultures have been used to understand basics of recombinant cell physiology, for process development and industrial production of recombinant proteins. As semi-continuous perfusion processes have been used for mammalian cell-based recombinant processes for a long time and reviewed elsewhere [ 10 , 11 , 12 ], our review focuses on microorganisms.…”

Section: Introductionmentioning

confidence: 99%

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Peebo

Neubauer

2018

Microorganisms

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Depending on the environmental conditions, cells adapt their metabolism and specific growth rate. Rearrangements occur on many different levels such as macromolecular composition, gene and protein expression, morphology and metabolic flux patterns. As the interplay of these processes also determines the output of a recombinant protein producing system, having control over specific growth rate of the culture is advantageous. Continuous culture methods were developed to grow cells in a constant environment and have been used for decades to study basic microbial physiology in a controlled and reproducible manner. Our review summarizes the uses of continuous cultures in cell physiology studies and process development, with a focus on recombinant protein-producing microorganisms.

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Section: Introductionmentioning

confidence: 99%

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Peebo

Neubauer

2018

Microorganisms

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“…20 Therefore, it is necessary to have dynamic experiments and high-frequency data to understand the behavior of a biological process. 21…”

Section: Introductionmentioning

confidence: 99%

Integrated Robotic Mini Bioreactor Platform for Automated, Parallel Microbial Cultivation With Online Data Handling and Process Control

et al. 2019

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“…Even though automated frameworks exist to harvest the culture at a desired biomass concentration [ 33 ], different growth rates, starting ODs and lag-phases make it difficult to reach the same OD at the same time when transforming different clones ( Figure 1 a). From the perspective of bioprocess engineering, the method of choice to maintain the cells at a given condition would be a continuous cultivation [ 34 ]. The most used system (due to its simplicity and robustness) is the chemostat, where the growth rate is determined by the dilution rate.…”

Section: Introductionmentioning

confidence: 99%

Automated Cell Treatment for Competence and Transformation of Escherichia coli in a High-Throughput Quasi-Turbidostat Using Microtiter Plates

et al. 2018

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Metabolic engineering and genome editing strategies often lead to large strain libraries of a bacterial host. Nevertheless, the generation of competent cells is the basis for transformation and subsequent screening of these strains. While preparation of competent cells is a standard procedure in flask cultivations, parallelization becomes a challenging task when working with larger libraries and liquid handling stations as transformation efficiency depends on a distinct physiological state of the cells. We present a robust method for the preparation of competent cells and their transformation. The strength of the method is that all cells on the plate can be maintained at a high growth rate until all cultures have reached a defined cell density regardless of growth rate and lag phase variabilities. This allows sufficient transformation in automated high throughput facilities and solves important scheduling issues in wet-lab library screenings. We address the problem of different growth rates, lag phases, and initial cell densities inspired by the characteristics of continuous cultures. The method functions on a fully automated liquid handling platform including all steps from the inoculation of the liquid cultures to plating and incubation on agar plates. The key advantage of the developed method is that it enables cell harvest in 96 well plates at a predefined time by keeping fast growing cells in the exponential phase as in turbidostat cultivations. This is done by a periodic monitoring of cell growth and a controlled dilution specific for each well. With the described methodology, we were able to transform different strains in parallel. The transformants produced can be picked and used in further automated screening experiments. This method offers the possibility to transform any combination of strain- and plasmid library in an automated high-throughput system, overcoming an important bottleneck in the high-throughput screening and the overall chain of bioprocess development.

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Continuous Bioprocess Development: Methods for Control and Characterization of the Biological System

Cited by 3 publications

References 111 publications

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Application of Continuous Culture Methods to Recombinant Protein Production in Microorganisms

Integrated Robotic Mini Bioreactor Platform for Automated, Parallel Microbial Cultivation With Online Data Handling and Process Control

Automated Cell Treatment for Competence and Transformation of Escherichia coli in a High-Throughput Quasi-Turbidostat Using Microtiter Plates

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