Evaluation of criteria for bioreactor comparison and operation standardization for mammalian cell culture

Barradas, Oscar Platas; Jandt, Uwe; Phan, Linh Da Minh; Villanueva, Mario E.; Schaletzky, Martin; Rath, Alexander; Freund, Susann; Reichl, Udo; Skerhutt, Eva; Scholz, Sebastian; Noll, Thomas; Sandig, Volker; Pörtner, Ralf; Zeng, An‐Ping

doi:10.1002/elsc.201100163

Cited by 35 publications

(18 citation statements)

References 30 publications

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“…Improvement in kinetic parameters observed for the experiments in bioreactor with respect to those from spinner are justified by the control of important variables for cell growth and metabolism, such as: pH, and dissolved oxygen concentration. Specifically, the maximum specific growth rate in bioreactor was two-fold higher than the values obtained for this parameter in spinner flasks (present work) and others, commonly reported for mammalian cell culture in bioreactors using serum free medium (Platas-Barradas et al 2012). Probably, fetal bovine serum addition to the culture medium justified this finding (Sidoli et al 2004).…”

Section: Discussionsupporting

confidence: 44%

Approach toward an efficient inoculum preparation stage for suspension BHK-21 cell culture

et al. 2014

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Mammalian cells are the most frequently used hosts for biopharmaceutical proteins manufacturing. Inoculum quality is a key element for establishing an efficient bioconversion process. The main objective in inoculation expansion process is to generate large volume of viable cells in the shortest time. The aim of this paper was to optimize the inoculum preparation stage of baby hamster kidney (BHK)-21 cells for suspension cultures in benchtop bioreactors, by means of a combination of static and agitated culture systems. Critical parameters for static (liquid column height: 5, 10, 15 mm) and agitated (working volume: 35, 50, 65 mL, inoculum volume percentage: 10, 30 % and agitation speed: 25, 60 rpm) cultures were study in T-flask and spinner flask, respectively. The optimal liquid column height was 5 mm for static culture. The maximum viable cell concentration in spinner flask cultures was reached with 50 mL working volume and the inoculum volume percentage was not significant in the range under study (10-30 %) at 25 rpm agitation. Agitation speed at 60 rpm did not change the main kinetic parameters with respect to those observed for 25 rpm. These results allowed for a schedule to produce more than 4 9 10 9 BHK-21 cells from 4 9 10 6 cells in 13 day with 1,051 mL culture medium.

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Section: Discussionsupporting

confidence: 44%

Approach toward an efficient inoculum preparation stage for suspension BHK-21 cell culture

et al. 2014

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“…The cultivation of synchronized CHO DP‐12 cells was performed in the bench top bioreactor Medorex Vario 1000 (Medorex, Germany) with a working volume of 100 ml. The temperature was controlled at 36.8°C, pH at 7.1 (

{CO}_{2}

or 0.5 M

{Na}_{2} {CO}_{3}

), agitation at 200 rpm (2.9 W/m 3 ; Platas Barradas et al, ) and dissolved oxygen at minimal 40% (sparging with pure oxygen if necessary). Samples were taken every 3 hr.…”

Section: Methodsmentioning

confidence: 99%

Model‐based identification of cell‐cycle‐dependent metabolism and putative autocrine effects in antibody producing CHO cell culture

Möller

Korte

Pörtner

et al. 2018

Biotech & Bioengineering

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The understanding of cell-cycle-dependent population heterogeneities in mammalian cell culture and their influence on production rates is still limited. Furthermore, metabolic regulations arising from self-expressed signaling factors (autocrine/autoinhibitory factors) have been postulated in the past, but no determination of such effects have been made so far for fast-growing production Chinese hamster ovary (CHO) cells in chemically defined media. In this study, a novel approach combining near-physiological treatment of cells (including synchronization), population resolved mechanistic modeling and statistical analysis was developed to identify population inhomogeneities. Cell-cycle-dependent population dynamics and metabolic regulations due to a putative autocrine factor were examined and their impact on the metabolic rates and antibody production of near-physiologically synchronized CHO DP-12 cell cultures was determined. To achieve this, a population resolved model was extended to describe putative autocrine-dependentt and cell-cycle-related metabolic rates for glucose, glutamine, lactate, ammonia, and antibody production. The model parameters were estimated based on data of two repeated batch cultivations (three batches each), with main substrates in excess and potentially inhibiting waste products (lactate and ammonium) controlled within narrow ranges. Significant changes, due to a putative autocrine factor, were identified for lactate and ammonia formation and antibody production. The cell growth and the uptake of glucose and glutamine were only partially affected by the putative autocrine under the given conditions. The results indicate the presence of a self-expressed autocrine factor and its strong impact on the metabolism of CHO DP-12 cells. Furthermore, glucose and glutamine uptake, as well as the formation of ammonium and antibody were found to be significantly cell-cycle-dependent. The combined approach has a strong potential to improve the understanding of the interplay of population heterogeneities and signal factors in mammalian cell culture.

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“…A variety of engineering criteria could be used for scale‐up . In mammalian cell cultures, one of the commonly used criteria is similar power input per volume ( P / V , or the mean specific energy dissipation rate). Other criteria, such as impeller pumping rate per unit bioreactor volume, and a combination of P / V with O 2 volumetric mass transfer coefficient (

k_{L} a true)

have been applied in stirred‐tank bioreactors with desirable scale‐up/process transfer performance.…”

Section: Introductionmentioning

confidence: 99%

A practical approach in bioreactor scale‐up and process transfer using a combination of constant P/V and vvm as the criterion

Hoshan

Jiang

et al. 2017

Biotechnology Progress

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Bioreactor scale-up is a critical step in the production of therapeutic proteins such as monoclonal antibodies (MAbs). With the scale-up criterion such as similar power input per volume or O volumetric mass transfer coefficient ( kLa), adequate oxygen supply and cell growth can be largely achieved. However, CO stripping in the growth phase is often inadequate. This could cascade down to increased base addition and osmolality, as well as residual lactate increase and compromised production and product quality. Here we describe a practical approach in bioreactor scale-up and process transfer, where bioreactor information may be limited. We evaluated the sparger kLa and kLaCO2 (CO volumetric mass transfer coefficient) from a range of bioreactor scales (3-2,000 L) with different spargers. Results demonstrated that kLa for oxygen is not an issue when scaling from small-scale to large-scale bioreactors at the same gas flow rate per reactor volume (vvm). Results also showed that sparging CO stripping, kLaCO2, is dominated by the gas throughput. As a result, a combination of a minimum constant vvm air or N flow with a similar specific power was used as the general scale-up criterion. An equation was developed to determine the minimum vvm required for removing CO produced from cell respiration. We demonstrated the effectiveness of using such scale-up criterion with five MAb projects exhibiting different cell growth and metabolic characteristics, scaled from 3 to 2,000 L bioreactors across four sites. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1146-1159, 2017.

show abstract

Evaluation of criteria for bioreactor comparison and operation standardization for mammalian cell culture

Cited by 35 publications

References 30 publications

Approach toward an efficient inoculum preparation stage for suspension BHK-21 cell culture

Approach toward an efficient inoculum preparation stage for suspension BHK-21 cell culture

Model‐based identification of cell‐cycle‐dependent metabolism and putative autocrine effects in antibody producing CHO cell culture

A practical approach in bioreactor scale‐up and process transfer using a combination of constant P/V and vvm as the criterion

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