Study of a recombinant CHO cell line producing a monoclonal antibody by ATF or TFF external filter perfusion in a WAVE Bioreactor™

Clincke, Marie-Françoise; Mölleryd, Carin; Zhang, Ye; Lindskog, Eva; Walsh, Kieron; Chotteau, Véronique

doi:10.1186/1753-6561-5-s8-p105

Cited by 35 publications

(20 citation statements)

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“…The spin-filter consists of a cylindrical screen, which is continuously rotating. The cell-free supernatant enters in the spin-filter from where it is pumped into the harvest line using TFF or ATF using polysulfone HF (Clincke et al 2011(Clincke et al , 2013a. They established steady state cultures at % 25 Â 10 6 cells/mL or 1-1.2 Â 10 8 cells/mL during more than 2 weeks and reached a maximal density of 2.14 Â 10 8 cells/mL.…”

Section: Tffmentioning

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: Tffmentioning

confidence: 99%

“…We performed a series of experiments in a wave bioreactor connected to an ATF2 using a MAb producing CHO-K1 DHFR À cell line (Clincke et al 2011(Clincke et al , 2013aChotteau et al 2014a). We developed and studied this system in several runs maintained at 20 Â 10 6 cells/mL by cell bleeding during up to 2 weeks.…”

Section: Atfmentioning

confidence: 99%

Perfusion Processes

Chotteau

2014

Cell Engineering

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“…The alternating tangential flow (ATF) system is a low shear filtration system that inhibits filter‐membrane fouling. It supports high cell densities, with moderate equipment cost . This external cell‐separation system is able to maintain continuous culture for extended periods of time and offers the capability of rapid filter change without compromising the culture run .…”

Section: Introductionmentioning

confidence: 99%

Alternating flow filtration as an alternative to internal spin filter based perfusion process: Impact on productivity and product quality

Bosco¹,

Paillet²,

Amadeo

et al. 2017

Biotechnology Progress

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This article reports the results obtained from comparison of internal spin filter (ISF) and alternating flow filtration (ATF) as cell retention systems, regarding cell growth, volumetric perfusion rate, cell specific perfusion rate and cell productivity in the fermentation process. As expected we were able to reach higher cell densities and to achieve longer runs since ATF systems are known to be less affected by fouling. Volumetric production of the reactor using the ATF system was 50-70% higher than the production achieved using the ISF due to higher cell density and a two-fold increase in the perfusion rate. On the other hand, downstream processing performances were evaluated regarding chromatographic steps yields and productivity and quality attributes of the purified materials. Similar results were obtained for all evaluated systems. The fact that we were able to achieve a 2 working volumes (WV)/day perfusion rate using an ATF system as cell retention device allowed us to virtually double the WV of a 25 L reactor. These results constitute valuable data for the optimization of recombinant protein production in perfusion processes since a two-fold increase in the average production of a manufacturing facility could be easily achieved as long as downstream scale up is possible. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1010-1014, 2017.

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“…Contrary to conventional filtration with cell broth moved in flow direction orthogonal to the filter surface, in TFF the cell suspension is flowing tangentially to the membrane potentially preventing membrane fouling . The ATF uses TFF but with the cell broth flow direction alternated using a diaphragm pump, creating less shear stress than the TFF peristaltic pump, and with a cycle time around 1 min . The alternated movement creates a back flush in the filter membrane, which can help preventing fouling…”

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

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201

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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Study of a recombinant CHO cell line producing a monoclonal antibody by ATF or TFF external filter perfusion in a WAVE Bioreactor™

Cited by 35 publications

References 0 publications

Perfusion Processes

Perfusion Processes

Alternating flow filtration as an alternative to internal spin filter based perfusion process: Impact on productivity and product quality

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

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