Batch‐to‐batch variability of two human designer cell lines – AGE1.HN and AGE1.HN.AAT – carried out by different laboratories under defined culture conditions using a mathematical model

Systems biology approaches involve collaboration of a larger number of research groups. Experiments are being performed in different laboratories dealing with different aspects of the topic of interest. Therefore, comparability of data collected for further analysis and modeling needs critical assessment. Here, growth and product formation of two human designer cell lines (AGE1.HN and its α1‐antitrypsin producing clone AGE1.HN.AAT) were investigated by four research laboratories. Cell lines were cultivated in shake flasks and stirred tank bioreactors operated at standardized conditions using a chemically defined medium, and a simple mathematical model was used to estimate characteristic process parameters. Results obtained for 35 batches showed that neither the initial viable cell concentration nor the initial concentration of glucose and glutamine showed significant differences between laboratories. For these measurements with low variations, specific growth rate and yields varied between 8.5 and 26% (relative standard error), indicating comparability of cultivations between laboratories for exponential growth. Higher variations of fitted parameters related to measurements with high initial variation. Comparing the nonproducing with the producing cell line, no significant differences were found regarding growth dynamics and metabolism. Overall, it seems justified to draw conclusions based on the entire experimental dataset of this systems biology project.

[1]  E. Heinzle,et al.  Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis , 2010, Bioprocess and biosystems engineering.

[2]  F. Wurm,et al.  Rational vector design and multi-pathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g/l by transient transfection under serum-free conditions , 2008, Nucleic acids research.

[3]  Rui Oliveira,et al.  Improving baculovirus production at high cell density through manipulation of energy metabolism. , 2010, Metabolic engineering.

[4]  H. Blanch,et al.  A kinetic analysis of hybridoma growth and metabolism in continuous suspension culture on serum‐free medium , 1991, Biotechnology and bioengineering.

[5]  P Lane,et al.  Improvement of a mammalian cell culture process by adaptive, model-based dialysis fed-batch cultivation and suppression of apoptosis , 2003, Bioprocess and biosystems engineering.

[6]  T. Schäfer,et al.  Modelling hybridoma cell growth and metabolism--a comparison of selected models and data. , 1996, Journal of biotechnology.

[7]  Julio R. Banga,et al.  Scatter search for chemical and bio-process optimization , 2007, J. Glob. Optim..

[8]  D. Kirwan,et al.  Effects of stirring and sparging on cultured hybridoma cells , 1990, Biotechnology and bioengineering.

[9]  Yih Yean Lee,et al.  Identification and expression analysis of miRNAs during batch culture of HEK-293 cells. , 2009, Journal of biotechnology.

[10]  D. Lütkemeyer,et al.  Optimization of serum-free fermentation processes for antibody production , 2004, Cytotechnology.

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

[12]  R. Linhardt,et al.  Engineering of routes to heparin and related polysaccharides , 2011, Applied Microbiology and Biotechnology.

[13]  Jose C. Menezes,et al.  Predicting mab product yields from cultivation media components, using near‐infrared and 2D‐fluorescence spectroscopies , 2011, Biotechnology progress.

[14]  M. Moo-young,et al.  Statistical methods in media optimization for batch and fed-batch animal cell culture , 2007, Bioprocess and biosystems engineering.

[15]  Joost D de Bruijn,et al.  Growth, metabolism, and growth inhibitors of mesenchymal stem cells. , 2009, Tissue engineering. Part A.

[16]  Udo Reichl,et al.  Segregated Mathematical Model for Growth of Anchorage‐Dependent MDCK Cells in Microcarrier Culture , 2008, Biotechnology progress.

[17]  F. Wurm.  Production of recombinant protein therapeutics in cultivated mammalian cells , 2004, Nature Biotechnology.

[18]  Olivier Henry,et al.  Enhanced glycoprotein production in HEK-293 cells expressing pyruvate carboxylase. , 2011, Metabolic engineering.

[19]  S. R. Nathan,et al.  A systematic approach to the validation of process control parameters for monoclonal antibody production in fed-batch culture of a murine myeloma. , 2000, Biotechnology and bioengineering.

[20]  T. Logtenberg,et al.  High‐Level Expression of Recombinant IgG in the Human Cell Line PER.C6 , 2003, Biotechnology progress.

[21]  J. Davies,et al.  Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: Expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII. , 2001, Biotechnology and bioengineering.

[22]  Robert V. Brill,et al.  Applied Statistics and Probability for Engineers , 2004, Technometrics.

[23]  Herbert Märkl,et al.  Effect of NH3 on the cell growth of a hybridoma cell line , 2004, Cytotechnology.

[24]  Mats Jirstrand,et al.  Systems biology Systems Biology Toolbox for MATLAB : a computational platform for research in systems biology , 2006 .

[25]  E. Heinzle,et al.  Effects of drugs in subtoxic concentrations on the metabolic fluxes in human hepatoma cell line Hep G2. , 2009, Toxicology and applied pharmacology.

[26]  Gary Walsh,et al.  Biopharmaceutical benchmarks 2006 , 2006, Nature Biotechnology.

[27]  J. Piret,et al.  Limitations to the amplification and stability of human tissue-type plasminogen activator expression by Chinese hamster ovary cells. , 2000, Biotechnology and bioengineering.

[28]  Gary Walsh,et al.  Biopharmaceutical benchmarks , 2000, Nature Biotechnology.

[29]  Zizhuo Xing,et al.  Modeling kinetics of a large‐scale fed‐batch CHO cell culture by Markov chain Monte Carlo method , 2010, Biotechnology progress.

[30]  J. M. Engasser,et al.  Determination of cell lysis and death kinetics in continuous hybridoma cultures from the measurement of lactate dehydrogenase release , 2004, Cytotechnology.

[31]  E. Heinzle,et al.  Metabolic flux rearrangement in the amino acid metabolism reduces ammonia stress in the α1-antitrypsin producing human AGE1.HN cell line. , 2012, Metabolic engineering.

[32]  C. Wittmann,et al.  In vivo quantification of intracellular amino acids and intermediates of the methionine pathway in Corynebacterium glutamicum. , 2005, Analytical biochemistry.

[33]  Bahar Bilgen,et al.  Characterization of mixing in a novel wavy-walled bioreactor for tissue engineering. , 2005, Biotechnology and bioengineering.

[34]  Udo Reichl,et al.  Evaluation of criteria for bioreactor comparison and operation standardization for mammalian cell culture , 2012 .

[35]  Raul H. C. Lopes,et al.  A two-dimensional Kolmogorov-Smirnov test , 2009 .

[36]  E. Heinzle,et al.  Primary metabolism in the new human cell line AGE1.HN at various substrate levels: increased metabolic efficiency and α1-antitrypsin production at reduced pyruvate load , 2011, Applied Microbiology and Biotechnology.

[37]  M. Yap,et al.  Enhancement of transient gene expression by fed-batch culture of HEK 293 EBNA1 cells in suspension , 2006, Biotechnology Letters.

[38]  F. Kudo,et al.  Differential effects of orbital and laminar shear stress on endothelial cells. , 2005, Journal of vascular surgery.

[39]  N. McElvaney,et al.  α1-Antitrypsin Deficiency: Biological Answers to Clinical Questions , 2001 .

[40]  W S Hu,et al.  Kinetic study of hybridoma cell growth in continuous culture: II. Behavior of producers and comparison to nonproducers , 1991, Biotechnology and bioengineering.

[41]  B O Palsson,et al.  Growth, Metabolic, and Antibody Production Kinetics of Hybridoma Cell Culture: 2. Effects of Serum Concentration, Dissolved Oxygen Concentration, and Medium pH in a Batch Reactor , 1991, Biotechnology progress.

[42]  T. Omasa,et al.  Enhanced antibody production following intermediate addition based on flux analysis in mammalian cell continuous culture , 2010, Bioprocess and biosystems engineering.