Application of bioreactor design principles and multivariate analysis for development of cell culture scale down models

A bench scale cell culture model representative of manufacturing scale (2,000 L) was developed based on oxygen mass transfer principles, for a CHO‐based process producing a recombinant human protein. Cell culture performance differences across scales are characterized most often by sub‐optimal performance in manufacturing scale bioreactors. By contrast in this study, reduced growth rates were observed at bench scale during the initial model development. Bioreactor models based on power per unit volume (P/V), volumetric mass transfer coefficient (kLa), and oxygen transfer rate (OTR) were evaluated to address this scale performance difference. Lower viable cell densities observed for the P/V model were attributed to higher sparge rates and reduced oxygen mass transfer efficiency (kLa) of the small scale hole spargers. Increasing the sparger kLa by decreasing the pore size resulted in a further decrease in growth at bench scale. Due to sensitivity of the cell line to gas sparge rate and bubble size that was revealed by the P/V and kLa models, an OTR model based on oxygen enrichment and increased P/V was selected that generated endpoint sparge rates representative of 2,000 L scale. This final bench scale model generated similar growth rates as manufacturing. In order to take into account other routinely monitored process parameters besides growth, a multivariate statistical approach was applied to demonstrate validity of the small scale model. After the model was selected based on univariate and multivariate analysis, product quality was generated and verified to fall within the 95% confidence limit of the multivariate model. Biotechnol. Bioeng. 2015;112: 84–97. © 2014 Wiley Periodicals, Inc.

[1]  Theodora Kourti,et al.  Process Analytical Technology Beyond Real-Time Analyzers: The Role of Multivariate Analysis , 2006 .

[2]  Hu Zhang,et al.  Engineering considerations for process development in mammalian cell cultivation. , 2010, Current pharmaceutical biotechnology.

[3]  A. Nienow.  Reactor Engineering in Large Scale Animal Cell Culture , 2006, Cytotechnology.

[4]  I. Karimi,et al.  Combined data preprocessing and multivariate statistical analysis characterizes fed-batch culture of mouse hybridoma cells for rational medium design. , 2010, Journal of biotechnology.

[5]  M. Callegari,et al.  Instrumental Robots Design with Applications to Manufacturing 7.1 Introduction the Design Cycle for Instrumental Robots 7.2 the Design of Function-oriented Robots Conceptual Design of Task-driven Robot-arms @bullet Conceptual Design of Work-constrained Robot-arms @bullet Computer Aids Based on Funct , 2001 .

[6]  Ulrich Eggers,et al.  Biochemical Engineering Fundamentals , 2016 .

[7]  Gail Sofer,et al.  Process Validation with a CMO , 2012 .

[8]  Mareike Harmsen,et al.  Development and fine-tuning of a scale down model for process characterization studies of a monoclonal antibody upstream production process , 2011, BMC proceedings.

[9]  A. Rathore,et al.  Quality by design for biopharmaceuticals , 2009, Nature Biotechnology.

[10]  Jennifer Gangi,et al.  Fed‐batch bioreactor process scale‐up from 3‐L to 2,500‐L scale for monoclonal antibody production from cell culture , 2007, Biotechnology and bioengineering.

[11]  W. Miller,et al.  Effects of CO2 and osmolality on hybridoma cells: growth, metabolism and monoclonal antibody production , 1998, Cytotechnology.

[12]  Ningning Ma,et al.  Quantitative Studies of Cell‐Bubble Interactions and Cell Damage at Different Pluronic F‐68 and Cell Concentrations , 2004, Biotechnology progress.

[13]  G. Karypis,et al.  Multivariate analysis of cell culture bioprocess data--lactate consumption as process indicator. , 2012, Journal of biotechnology.

[14]  Dale E Seborg,et al.  Fault Detection and Diagnosis in an Industrial Fed‐Batch Cell Culture Process , 2007, Biotechnology progress.

[15]  Sigma S Mostafa,et al.  Strategies for Improved dCO2 Removal in Large‐Scale Fed‐Batch Cultures , 2003, Biotechnology progress.

[16]  Gail Sofer,et al.  Leveraging Multivariate Analysis Tools to Qualify Scaled- Down Models , 2012 .

[17]  Jeremy S. Conner,et al.  Application of Multivariate Analysis toward Biotech Processes: Case Study of a Cell‐Culture Unit Operation , 2007, Biotechnology progress.

[18]  Thomas Ryll,et al.  Maximizing productivity of CHO cell‐based fed‐batch culture using chemically defined media conditions and typical manufacturing equipment , 2010, Biotechnology progress.

[19]  Graham McCreath,et al.  Practical Considerations for DoE Implementation in Quality By Design , 2010 .

[20]  C. Goochee,et al.  Structural Features of Nonionic Polyglycol Polymer Molecules Responsible for the Protective Effect in Sparged Animal Cell Bioreactors , 1990, Biotechnology progress.

[21]  Anurag S. Rathore,et al.  Scaling down of biopharmaceutical unit operations: Part I: Fermentation , 2005 .

[22]  George G. Chase,et al.  Improved scale-up strategies of bioreactors , 1992 .

[23]  Weichang Zhou,et al.  NS0 cell damage by high gas velocity sparging in protein‐free and cholesterol‐free cultures , 2008, Biotechnology and bioengineering.

[24]  A. Rathore,et al.  Application of Multivariate Data Analysis for Identification and Successful Resolution of a Root Cause for a Bioprocessing Application , 2008, Biotechnology progress.

[25]  D. Inlow,et al.  CO2 in large-scale and high-density CHO cell perfusion culture , 2004, Cytotechnology.

[26]  S J Meier,et al.  Cell death from bursting bubbles: role of cell attachment to rising bubbles in sparged reactors. , 1999, Biotechnology and bioengineering.

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

[28]  Thomas Ryll,et al.  Development of a scale down cell culture model using multivariate analysis as a qualification tool , 2014, Biotechnology progress.

[29]  Zizhuo Xing,et al.  Scale‐up analysis for a CHO cell culture process in large‐scale bioreactors , 2009, Biotechnology and bioengineering.

[30]  Hong Sun,et al.  Development of a scaled-down aerobic fermentation model for scale-up in recombinant protein vaccine manufacturing. , 2012, Vaccine.

[31]  John Pieracci,et al.  Leveraging Multivariate Analysis Tools to Qualify Scaled-Down Models , 2012 .

[32]  Advanced methods for bioreactor characterization. , 1992, Journal of biotechnology.

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

[34]  David E Block,et al.  An integrated approach to optimization of Escherichia coli fermentations using historical data , 2003, Biotechnology and bioengineering.

[35]  Eric N M van Sprang,et al.  Multivariate data analysis on historical IPV production data for better process understanding and future improvements , 2010, Biotechnology and bioengineering.

[36]  Feng Li,et al.  A Systematic Approach for Scale‐Down Model Development and Characterization of Commercial Cell Culture Processes , 2006, Biotechnology progress.

[37]  J Tramper,et al.  Lethal events during gas sparging in animal cell culture , 1991, Biotechnology and bioengineering.

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

[39]  M. Moo-young,et al.  Gas holdup in pneumatic reactors , 1988 .