Development of a Large Scale Process for the Production of Recombinant Truncated Factor VIII in CHO Cells under Cell Growth Arrest Conditions

Chotteau, Véronique; Björling, Torsten; Boork, Sarah; Brink-Nilsson, H; Chatzissavidou, Nathalie; Fenge, Christel; Lindner-Ollsson, Elisabeth; Olofsson, Mats; Rosenquist, Johan; Sandberg, Helena; Smeds, Anna‐Lisa; Drapeau, Denis

doi:10.1007/978-94-010-0369-8_67

Cited by 4 publications

(6 citation statements)

References 5 publications

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“…Cell arrest obtained by hypothermia or addition of a chemical, e.g. butyrate, has been reported for perfusion; given that the cell specific productivity is comparable or increased and that the protein quality is correct or improved (Ducommun et al 2002a;Chotteau et al 2001;Chen et al 2004;Kim et al 2004;Angepat et al 2005;Oh et al 2005;Meuwly et al 2006;Ahn et al 2008;Zhang et al 2014a). Another advantage of lowering the temperature under 35.5 C is that the proteolytic activity is lower compared to 37 C (Chuppa et al 1997).…”

Section: Cell Arrestmentioning

confidence: 93%

“…A short residence time is more favourable for the product quality and is mandatory for the labile glycoprotein, e.g. factor VIII, enzymes, (Chuppa et al 1997;Sandberg et al 2006;Chotteau et al 2001). Thanks to the higher volumetric production and longer cultivation length, bioreactors smaller than the ones used for fed-batch processes by at least one order of magnitude can be used to manufacture equivalent amounts of the product of interest (POI).…”

Section: Introductionmentioning

confidence: 99%

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Perfusion Processes

Chotteau

2014

Cell Engineering

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The interest for perfusion is increasing nowadays. This new focus has emerged from a synergy of a demand for disposable equipment and the availability of robust cell separation device, as well as the need for higher flexibility and lower investment cost. The cell separation devices mostly used today are based on filtration, i.e. alternating flow filtration, tangential flow filtration, spin-filter, or acceleration/gravity, i.e. inclined settler, centrifuge, acoustic settler. This paper gives an introduction to the basic concepts of perfusion and its practical implementation. It reviews the actual cell separation devices and describes the approaches used in the field to develop and optimize the perfusion processes.

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Section: Cell Arrestmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Perfusion Processes

Chotteau

2014

Cell Engineering

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“…In comparison to batch and fed‐batch modes, the perfusion mode allows prolonging healthy cultures, potentially at high cell density, as well as a short residence time of the product in the bioreactor. This is more favorable for the product quality and is mandatory for the production of unstable glycoprotein, for example, factor VIII . Another advantage of the perfusion mode is the use of smaller bioreactors compared with fed‐batch processes, which implies benefits such as reduced clean‐in‐place operation and the possibility to use disposable bioreactors instead of stainless steel reactors due to the smaller working volumes.…”

Section: Introductionmentioning

confidence: 99%

“…This is more favorable for the product quality and is mandatory for the production of unstable glycoprotein, for example, factor VIII. 6 Another advantage of the perfusion mode is the use of smaller bioreactors compared with fed-batch processes, which implies benefits such as reduced clean-in-place operation and the possibility to use disposable bioreactors instead of stainless steel reactors due to the smaller working volumes. Besides its use in biopharmaceutical production bioreactor, perfusion mode can also be used for high cell density seed bioreactors, cell bank manufacturing, or the production of proteins as research tool.…”

Section: Introductionmentioning

confidence: 99%

Very high density of CHO cells in perfusion by ATF or TFF in WAVE bioreactor™. Part I. Effect of the cell density on the process

Clincke

Mölleryd

Zhang

et al. 2013

Biotechnology Progress

202

138

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High cell density perfusion process of antibody producing CHO cells was developed in disposable WAVE Bioreactor™ using external hollow fiber filter as cell separation device. Both “classical” tangential flow filtration (TFF) and alternating tangential flow system (ATF) equipment were used and compared. Consistency of both TFF- and ATF-based cultures was shown at 20–35 × 106 cells/mL density stabilized by cell bleeds. To minimize the nutrients deprivation and by-product accumulation, a perfusion rate correlated to the cell density was applied. The cells were maintained by cell bleeds at density 0.9–1.3 × 108 cells/mL in growing state and at high viability for more than 2 weeks. Finally, with the present settings, maximal cell densities of 2.14 × 108 cells/mL, achieved for the first time in a wave-induced bioreactor, and 1.32 × 108 cells/mL were reached using TFF and ATF systems, respectively. Using TFF, the cell density was limited by the membrane capacity for the encountered high viscosity and by the pCO2 level. Using ATF, the cell density was limited by the vacuum capacity failing to pull the highly viscous fluid. Thus, the TFF system allowed reaching higher cell densities. The TFF inlet pressure was highly correlated to the viscosity leading to the development of a model of this pressure, which is a useful tool for hollow fiber design of TFF and ATF. At very high cell density, the viscosity introduced physical limitations. This led us to recommend cell densities under 1.46 × 108 cell/mL based on the analysis of the theoretical distance between the cells for the present cell line. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:754–767, 2013

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“…An optimized perfusion process using a sufficiently genetically stable cell line can potentially operate for months, and in some cases may be the only viable process for a labile molecule. [13][14][15] Furthermore, a continuous flow of a cell-free harvest stream exiting the bioreactor can be directly integrated with downstream unit operations providing additional advantages. A continuous bioprocess consolidates unit operations as fewer hold steps are needed and corresponding hold tanks are eliminated, also potentially reducing product quality issues associated with unintended degradation pathways.…”

Section: Introductionmentioning

confidence: 99%

Shift to high‐intensity, low‐volume perfusion cell culture enabling a continuous, integrated bioprocess

Gagnon

Nagre

Wang

et al. 2018

Biotechnology Progress

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In order to address the increasing demand for biologics, cell culture intensification using perfusion offers significantly higher productivities while also reducing manufacturing costs, especially when part of an integrated, continuous bioprocess. An initial study of a long‐duration perfusion process using a cell‐bleed to maintain a target cell density observed a 2.1‐fold higher cell‐specific productivity and a gradual decline in the culture growth rate when perfused at an overall lower rate. Subsequent studies sought an alternative process that largely reduced the overall volume of media needed by first perfusing at a high cell‐specific perfusion rate (CSPR) to support a high cell density followed by continued perfusion at a low CSPR to promote a more productive stationary phase. This high intensity, low‐volume perfusion (HILVOP) process achieved cumulative volumetric productivities of 1.5–1.6 g/L/day with two CHO cell lines. When compared to each cell line's respective commercial‐ready, fed‐batch process, a 3.1–3.8‐fold productivity increase was demonstrated while yielding similar product quality. Furthermore, the higher productivity achieved with HILVOP used 6.6–12.3‐fold less media than a similarly productive long‐duration process. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1472–1481, 2018

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Development of a Large Scale Process for the Production of Recombinant Truncated Factor VIII in CHO Cells under Cell Growth Arrest Conditions

Cited by 4 publications

References 5 publications

Perfusion Processes

Perfusion Processes

Very high density of CHO cells in perfusion by ATF or TFF in WAVE bioreactor™. Part I. Effect of the cell density on the process

Shift to high‐intensity, low‐volume perfusion cell culture enabling a continuous, integrated bioprocess

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