Characterization and comparison of ATF and TFF in stirred bioreactors for continuous mammalian cell culture processes

Abstract In this work, two external filtration devices for cell retention, the commercial alternating tangential flow (ATF) system and a tangential flow filtration (TFF) setup driven by a bearingless centrifugal pump were compared. Physical characterization of the bioreactor hydrodynamics revealed significantly smaller maximum stress values in the ATF at similar mixing and oxygen mass transfer rates. Suitable operating parameters in terms of aeration, stirring and operation of the retention device were determined. Steady culture of Chinese hamster ovary (CHO) cells at 20, 40 and 60 × 10 6 cells/mL for at least one week was achieved in perfusion at a fixed harvest rate of one reactor volume per day for both setups. Contrary to the ATF, considerable retention (up to 50%) of the produced monoclonal antibody in the TFF was observed, however without significant difference in product quality. The integration of physical and cell culture characterization allows the comprehensive assessment of cell culture performance. The presented approach can serve as a general procedure to develop a scalable mammalian cell perfusion culture.

[1]  K. Riet,et al.  Review of Measuring Methods and Results in Nonviscous Gas-Liquid Mass Transfer in Stirred Vessels , 1979 .

[2]  Paul F. Greenfield,et al.  Experience in scale-up of homogeneous perfusion culture for hybridomas , 1991 .

[3]  Matthias Reuss,et al.  Structured Modelling of Bioreactors , 1994 .

[4]  Véronique Chotteau,et al.  Very High Density of Chinese Hamster Ovary Cells in Perfusion by Alternating Tangential Flow or Tangential Flow Filtration in WAVE Bioreactor™—Part II: Applications for Antibody Production and Cryopreservation , 2013, Biotechnology progress.

[5]  R. Gentz,et al.  High-density perfusion culture of insect cells with a biosep ultrasonic filter. , 1998, Biotechnology and bioengineering.

[6]  R. Spier,et al.  A comparison of oxygenation methods fro high‐density perfusion culture of animal cells , 1993, Biotechnology and bioengineering.

[7]  Thomas K. Villiger,et al.  Experimental determination of maximum effective hydrodynamic stress in multiphase flow using shear sensitive aggregates , 2015 .

[8]  John Thrift,et al.  The "push-to-low" approach for optimization of high-density perfusion cultures of animal cells. , 2006, Advances in biochemical engineering/biotechnology.

[9]  Regine Eibl,et al.  Investigations on Mechanical Stress Caused to CHO Suspension Cells by Standard and Single‐Use Pumps , 2013 .

[10]  C. Goudar,et al.  Recent advances in the understanding of biological implications and modulation methodologies of monoclonal antibody N‐linked high mannose glycans , 2014, Biotechnology and bioengineering.

[11]  S E Builder,et al.  Industrial scale harvest of proteins from mammalian cell culture by tangential flow filtration , 1991, Biotechnology and bioengineering.

[12]  H. Chang,et al.  Limited Use of Centritech Lab II Centrifuge in Perfusion Culture of rCHO Cells for the Production of Recombinant Antibody , 2008, Biotechnology progress.

[13]  Ye Zhang,et al.  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 , 2013, Biotechnology progress.

[14]  James M Piret,et al.  Estimating cell specific oxygen uptake and carbon dioxide production rates for mammalian cells in perfusion culture , 2011, Biotechnology progress.

[15]  R. van Reis,et al.  Linear scale ultrafiltration. , 1997, Biotechnology and bioengineering.

[16]  D S Kompala,et al.  Inclined Sedimentation for Selective Retention of Viable Hybridomas in a Continuous Suspension Bioreactor , 1990, Biotechnology progress.

[17]  B. Maiorella,et al.  Crossflow microfiltration of animal cells , 1991, Biotechnology and bioengineering.

[18]  James M Piret,et al.  Scale‐up and optimization of an acoustic filter for 200 L/day perfusion of a CHO cell culture , 2002, Biotechnology and bioengineering.

[19]  C. Choo,et al.  High‐Level Production of a Monoclonal Antibody in Murine Myeloma Cells by Perfusion Culture Using a Gravity Settler , 2007, Biotechnology progress.

[20]  Thomas Ryll,et al.  Development and implementation of a perfusion‐based high cell density cell banking process , 2011, Biotechnology progress.

[21]  D D Ryu,et al.  Monoclonal antibody productivity and the metabolic pattern of perfusion cultures under varying oxygen tensions , 1993, Biotechnology and bioengineering.

[22]  D. Lütkemeyer,et al.  Mapping and partial characterization of proteases expressed by a CHO production cell line , 2006, Biotechnology and bioengineering.

[23]  D. Kompala,et al.  Production of a Secreted Glycoprotein from an Inducible Promoter System in a Perfusion Bioreactor , 2004, Biotechnology progress.

[24]  Hervé Broly,et al.  Tailoring recombinant protein quality by rational media design , 2015, Biotechnology progress.

[25]  Daniel Cummings,et al.  Integrated continuous production of recombinant therapeutic proteins , 2012, Biotechnology and bioengineering.

[26]  Massimo Morbidelli,et al.  Adaptation for survival: phenotype and transcriptome response of CHO cells to elevated stress induced by agitation and sparging. , 2014, Journal of biotechnology.

[27]  Thomas K. Villiger,et al.  Evaluating the impact of cell culture process parameters on monoclonal antibody N-glycosylation. , 2014, Journal of biotechnology.

[28]  J. Chalmers,et al.  Physiological responses of CHO cells to repetitive hydrodynamic stress , 2009, Biotechnology and bioengineering.

[29]  Angela Meier,et al.  Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns , 2013, Biotechnology and bioengineering.

[30]  W. L. Ling,et al.  Case Study: An accelerated 8‐day monoclonal antibody production process based on high seeding densities , 2013, Biotechnology progress.

[31]  K Konstantinov,et al.  Fermentor temperature as a tool for control of high-density perfusion cultures of mammalian cells. , 1997, Biotechnology and bioengineering.

[32]  Suzanne S Farid,et al.  Fed‐batch and perfusion culture processes: Economic, environmental, and operational feasibility under uncertainty , 2013, Biotechnology and bioengineering.

[33]  Franco Magelli,et al.  Analysis of KLa Measurement Methods in Stirred Vessels: The Role of Experimental Techniques and Fluid Dynamic Models , 2010 .

[34]  Sadettin S. Ozturk,et al.  Engineering challenges in high density cell culture systems , 2004, Cytotechnology.

[35]  H. Ziehr,et al.  Efficiency improvement of an antibody production process by increasing the inoculum density , 2014, Biotechnology progress.

[36]  D. Kompala,et al.  Viable Cell Recycle with an Inclined Settler in the Perfusion Culture of Suspended Recombinant Chinese Hamster Ovary Cells , 1994, Biotechnology progress.

[37]  C. Moresoli,et al.  Effect of pore size, shear rate, and harvest time during the constant permeate flux microfiltration of CHO cell culture supernatant , 2008, Biotechnology progress.

[38]  Z. Li,et al.  Optimal and consistent protein glycosylation in mammalian cell culture. , 2009, Glycobiology.

[39]  Jeffrey J. Chalmers,et al.  The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding , 2011, Cytotechnology.

[40]  Marco Jenzsch,et al.  Optimizing capacity utilization by large scale 3000 L perfusion in seed train bioreactors , 2013, Biotechnology progress.

[41]  D. Hatton,et al.  Determination of Chinese hamster ovary cell line stability and recombinant antibody expression during long‐term culture , 2012, Biotechnology and bioengineering.

[42]  A. R. Raspolli Galletti,et al.  An Innovative Microwave Process for Nanocatalyst Synthesis , 2010 .

[43]  Amine Kamen,et al.  Acoustic cell filter: a proven cell retention technology for perfusion of animal cell cultures. , 2004, Biotechnology advances.

[44]  Marcella Yu,et al.  Effects of cell culture conditions on antibody N‐linked glycosylation—what affects high mannose 5 glycoform , 2011, Biotechnology and bioengineering.

[45]  Jason Condon,et al.  Understanding and modeling alternating tangential flow filtration for perfusion cell culture , 2014, Biotechnology progress.

[46]  Massimo Morbidelli,et al.  Development of a Scale-Down Model of hydrodynamic stress to study the performance of an industrial CHO cell line under simulated production scale bioreactor conditions. , 2013, Journal of biotechnology.

[47]  Alvin W. Nienow,et al.  On impeller circulation and mixing effectiveness in the turbulent flow regime , 1997 .

[48]  M B Sliwkowski,et al.  Ammonium alters N-glycan structures of recombinant TNFR-IgG: degradative versus biosynthetic mechanisms. , 2000, Biotechnology and bioengineering.

[49]  A A Kamen,et al.  Use of the Centritech Lab Centrifuge for Perfusion Culture of Hybridoma Cells in Protein‐Free Medium , 1996, Biotechnology progress.

[50]  Massimo Morbidelli,et al.  Determination of the maximum operating range of hydrodynamic stress in mammalian cell culture. , 2015, Journal of biotechnology.