Investigations on Mechanical Stress Caused to CHO Suspension Cells by Standard and Single‐Use Pumps

Blaschczok, Katharina; Kaiser, Stefan; Löffelholz, Christian; Imseng, Nicole; Burkart, Jürg; Bösch, Pascal; Dornfeld, Wolfgang; Eibl, Regine; Eibl, Dieter

doi:10.1002/cite.201200135

Cited by 26 publications

(34 citation statements)

References 26 publications

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“…The impact of using a manifold and peristaltic pumps for cell culture broth recirculation is observed on Figure b, where an initial viability decay is observed on the Day 3 corresponding to the media exchange start. This observation is in‐line with the Blaschczok et al () study on the impact of peristaltic pumps on cell culture viability. The increased mechanical stress by the peristaltic pump leads to the release of intracellular components and an increase of cell debris creating a more challenging filtration scenario.…”

Section: Resultssupporting

confidence: 90%

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Impact of micro and macroporous TFF membranes on product sieving and chromatography loading for perfusion cell culture

Pinto

Napoli

Brower

2019

Biotech & Bioengineering

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Bioprocess intensification can be achieved through high cell density perfusion cell culture with continuous protein capture integration. Protein passage and cell retention are commonly accomplished using tangential flow filtration systems consisting of microporous membranes. Significant challenges, including low efficiency and decaying product sieving over time, are commonly observed in these cell retention devices. Here, we demonstrate that a macroporous membrane overcomes the product sieving challenges when comparing to several other membrane chemistries and pore sizes within the microporous range. This way, variable chromatography column loading is avoided. The macroporous membrane yielded a 13,000 L/m2 volumetric throughput. The membrane's cut‐off size results in an increased permeate turbidity due to particles passage, such as cell debris, through pores ranging from 1 to 4 µm. In addition, successful chromatography column plugging mitigation was achieved by employing depth filtration before the chromatographic step. Depth filtration volumetric throughputs were between 600 and 1,000 L/m2. Combing a macroporous cell retention device with a depth filter not only provided an alternative to address the challenge of undesired long protein residence times in the bioreactor due to product sieving decay, but also exhibited a throughput increase, making the integration of multicolumn capture chromatography with a perfusion cell culture a more robust process.

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Section: Resultssupporting

confidence: 90%

“…This observation is in-line with the Blaschczok et al (2013) study on the impact of peristaltic pumps on cell culture viability. The increased mechanical stress by the peristaltic pump leads to the release of intracellular components and an increase of cell debris creating a more challenging filtration scenario.…”

Section: Tff Membrane Screeningsupporting

confidence: 89%

Impact of micro and macroporous TFF membranes on product sieving and chromatography loading for perfusion cell culture

Pinto

Napoli

Brower

2019

Biotech & Bioengineering

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“…However, the use of peristaltic pumps can lead to significant hydrodynamic damage, affecting cell growth and cell viability, which is furthermore strongly dependent on the specific cell line and clone [12,43]. Therefore, other circulation systems as syringe pumps or centrifugal pumps might be more suitable for the application in scale-down models [12,44]. However, batch fermentations including a recirculation loop and peristaltic pumps revealed no effects of the circulation system on the applied cell line in this study (data not shown) and thus the circulation system was not considered to be the root cause for the decreased cell growth in the two-compartment system.…”

Section: Evaluation Of Growth Characteristics Of the Twocompartment Smentioning

confidence: 99%

The impact of pH inhomogeneities on CHO cell physiology and fed‐batch process performance – two‐compartment scale‐down modelling and intracellular pH excursion

Brunner

Braun

Doppler

et al. 2017

Biotechnology Journal

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Due to high mixing times and base addition from top of the vessel, pH inhomogeneities are most likely to occur during large-scale mammalian processes. The goal of this study was to set-up a scale-down model of a 10-12 m stirred tank bioreactor and to investigate the effect of pH perturbations on CHO cell physiology and process performance. Short-term changes in extracellular pH are hypothesized to affect intracellular pH and thus cell physiology. Therefore, batch fermentations, including pH shifts to 9.0 and 7.8, in regular one-compartment systems are conducted. The short-term adaption of the cells intracellular pH are showed an immediate increase due to elevated extracellular pH. With this basis of fundamental knowledge, a two-compartment system is established which is capable of simulating defined pH inhomogeneities. In contrast to state-of-the-art literature, the scale-down model is included parameters (e.g. volume of the inhomogeneous zone) as they might occur during large-scale processes. pH inhomogeneity studies in the two-compartment system are performed with simulation of temporary pH zones of pH 9.0. The specific growth rate especially during the exponential growth phase is strongly affected resulting in a decreased maximum viable cell density and final product titer. The gathered results indicate that even short-term exposure of cells to elevated pH values during large-scale processes can affect cell physiology and overall process performance. In particular, it could be shown for the first time that pH perturbations, which might occur during the early process phase, have to be considered in scale-down models of mammalian processes.

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“…During the pumping process, mechanical stress can result in irreversible cell damage, which might impair the quantity and quality of the product [1][2][3][4] . The level of mechanical stress depends on the pump type and pump settings, as demonstrated in previous studies [5][6] . Commonly, peristaltic, syringe and diaphragm pumps are used for single-use (SU) technology based applications.…”

Section: Introductionmentioning

confidence: 94%

“…The motor is magnetically driven in order to avoid narrow gaps between the impeller and the pump housing (Figure 1). A previous study investigated the MagLev centrifugal pumps and showed lower mechanical stress in Chinese Hamster Ovary (CHO) cells compared with peristaltic and 4-piston diaphragm pumps 5 . In addition, hemolysis analyses revealed no significant blood trauma and thrombus formation over a range of operation conditions using these pumps [8][9][10][11] .…”

Section: Introductionmentioning

confidence: 99%

A Cost-effective and Reliable Method to Predict Mechanical Stress in Single-use and Standard Pumps

Dittler

Dornfeld

Schöb

et al. 2015

JoVE

Self Cite

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Pumps are mainly used when transferring sterile culture broths in biopharmaceutical and biotechnological production processes. However, during the pumping process shear forces occur which can lead to qualitative and/or quantitative product loss. To calculate the mechanical stress with limited experimental expense, an oil-water emulsion system was used, whose suitability was demonstrated for drop size detections in bioreactors 1 . As drop breakup of the oil-water emulsion system is a function of mechanical stress, drop sizes need to be counted over the experimental time of shear stress investigations. In previous studies, the inline endoscopy has been shown to be an accurate and reliable measurement technique for drop size detections in liquid/liquid dispersions. The aim of this protocol is to show the suitability of the inline endoscopy technique for drop size measurements in pumping processes. In order to express the drop size, the Sauter mean diameter d 32 was used as the representative diameter of drops in the oil-water emulsion. The results showed low variation in the Sauter mean diameters, which were quantified by standard deviations of below 15%, indicating the reliability of the measurement technique.

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Investigations on Mechanical Stress Caused to CHO Suspension Cells by Standard and Single‐Use Pumps

Cited by 26 publications

References 26 publications

Impact of micro and macroporous TFF membranes on product sieving and chromatography loading for perfusion cell culture

Impact of micro and macroporous TFF membranes on product sieving and chromatography loading for perfusion cell culture

The impact of pH inhomogeneities on CHO cell physiology and fed‐batch process performance – two‐compartment scale‐down modelling and intracellular pH excursion

A Cost-effective and Reliable Method to Predict Mechanical Stress in Single-use and Standard Pumps

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