Application of bioreactor design principles and multivariate analysis for development of cell culture scale down models
Abstract
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 (kL a), 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 (kL a) of the small scale hole spargers. Increasing the sparger kL a 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 kL a 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.
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The approach used can serve as a model development procedure for cell culture processes.
“the approach used can serve as a model development procedure for cell culture processes.”
From Factors Affecting Microalgae Production for Biofuels and the Potentials of Chemometric Methods in Assessing and Optimizing Productivity (2019) · cites this paper as [79]
Tescoine et al. applied principal component analysis in the design of a cell culture bioreactor.
“Tescoine et al. [79] applied the principal component analysis (PCA) in the design of a cell culture bioreactor.”
From Factors Affecting Microalgae Production for Biofuels and the Potentials of Chemometric Methods in Assessing and Optimizing Productivity (2019) · cites this paper as [79]
The PCA module was built using four principal components that represented 85% of the initial data set.
“PCA module was built using four principal components that represented 85% of the initial data set.”
From Factors Affecting Microalgae Production for Biofuels and the Potentials of Chemometric Methods in Assessing and Optimizing Productivity (2019) · cites this paper as [79]
Product quality was generated and verified following the model, and the verified results fell within the 95% confidence limit of the model.
“Subsequently, the product quality was generated and verified following the model. The verified results fell within 95% confidence limit of the model”
From Factors Affecting Microalgae Production for Biofuels and the Potentials of Chemometric Methods in Assessing and Optimizing Productivity (2019) · cites this paper as [79]
Cites
Biochemical engineering fundamentals
Peppas, N. A. (1986) · Journal of Controlled Release · doi:10.1016/0168-3659(86)90022-2 · Cited by 2,350 · see citing works
Quality by design for biopharmaceuticals
Rathore, A., & Winkle, H. (2009) · Nature Biotechnology · doi:10.1038/nbt0109-26 · Cited by 891 · see citing works
Review of Measuring Methods and Results in Nonviscous Gas-Liquid Mass Transfer in Stirred Vessels
Riet, K. V. (1979) · Industrial & Engineering Chemistry Process Design and Development · doi:10.1021/i260071a001 · Cited by 770 · see citing works
Quality by Design in Biopharmaceuticals
Yang, H. (2016) · doi:10.1201/9781315371726-7 · Cited by 545 · see citing works
Reactor Engineering in Large Scale Animal Cell Culture
Nienow, A. W. (2006) · Cytotechnology · doi:10.1007/s10616-006-9005-8 · Cited by 433 · see citing works
Huang, Y., Hu, W., Rustandi, E., Chang, K., Yusuf‐Makagiansar, H., & Ryll, T. (2010) · Biotechnology Progress · doi:10.1002/btpr.436 · Cited by 353 · see citing works
Self-citationScale‐up analysis for a CHO cell culture process in large‐scale bioreactors
Xing, Z., Kenty, B. M., Li, Z. J., & Lee, S. S. (2009) · Biotechnology and Bioengineering · doi:10.1002/bit.22287 · Cited by 228 · see citing works
Engineering challenges in high density cell culture systems
Ozturk, S. S. (1996) · Cytotechnology · doi:10.1007/bf00353919 · Cited by 201 · see citing works
Multivariate analysis of cell culture bioprocess data—Lactate consumption as process indicator
Le, H., Kabbur, S., Pollastrini, L., Sun, Z., Mills, K., Johnson, K., Karypis, G., & Hu, W.-S. (2012) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2012.08.021 · Cited by 177 · see citing works
CO2 in large-scale and high-density CHO cell perfusion culture
Gray, D. R., Chen, S., Howarth, W., Inlow, D., & Maiorella, B. L. (1996) · Cytotechnology · doi:10.1007/bf00353925 · Cited by 176 · see citing works
The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding
Hu, W., Berdugo, C., & Chalmers, J. J. (2011) · Cytotechnology · doi:10.1007/s10616-011-9368-3 · Cited by 150 · see citing works
Lethal events during gas sparging in animal cell culture
Jöbses, I. M. L., Martens, D. E., & TRAMPER, J. (1991) · Biotechnology and Bioengineering · doi:10.1002/bit.260370510 · Cited by 119 · see citing works
Murhammer, D. W., & Goochee, C. F. (1990) · Biotechnology Progress · doi:10.1021/bp00002a008 · Cited by 114 · see citing works
Strategies for Improved dCO <sub>2</sub> Removal in Large‐Scale Fed‐Batch Cultures
Mostafa, S. S., & Gu, X. (. (2003) · Biotechnology Progress · doi:10.1021/bp0256263 · Cited by 113 · see citing works
Improved scale-up strategies of bioreactors
Ju, L., & Chase, G. G. (1992) · Bioprocess and Biosystems Engineering · doi:10.1007/bf00369263 · Cited by 112 · see citing works
Li, F., Hashimura, Y., Pendleton, R., Harms, J., Collins, E., & Lee, B. (2006) · Biotechnology Progress · doi:10.1021/bp0504041 · Cited by 110 · see citing works
Yang, J., Lu, C., Stasny, B., Henley, J., Guinto, W., Gonzalez, C., Gleason, J., Fung, M., Collopy, B., Benjamino, M., et al. (2007) · Biotechnology and Bioengineering · doi:10.1002/bit.21413 · Cited by 100 · see citing works
Kirdar, A. O., Conner, J. S., Baclaski, J., & Rathore, A. S. (2007) · Biotechnology Progress · doi:10.1021/bp060377u · Cited by 96 · see citing works
Kirdar, A. O., Green, K. D., & Rathore, A. S. (2008) · Biotechnology Progress · doi:10.1021/bp0704384 · Cited by 95 · see citing works
Ma, N., Chalmers, J. J., Aunins, J. G., Zhou, W., & Xie, L. (2004) · Biotechnology Progress · doi:10.1021/bp0342405 · Cited by 86 · see citing works
deZengotita, V. M., Kimura, R., & Miller, W. M. (1998) · Cytotechnology · doi:10.1023/a:1008010605287 · Cited by 78 · see citing works
Process Analytical Technology Beyond Real-Time Analyzers: The Role of Multivariate Analysis
Kourti, T. (2006) · Critical Reviews in Analytical Chemistry · doi:10.1080/10408340600969957 · Cited by 74 · see citing works
Development of a scale down cell culture model using multivariate analysis as a qualification tool
Tsang, V. L., Wang, A. X., Yusuf‐Makagiansar, H., & Ryll, T. (2013) · Biotechnology Progress · doi:10.1002/btpr.1819 · Cited by 64 · see citing works
Self-citationFault Detection and Diagnosis in an Industrial Fed-Batch Cell Culture Process
Gunther, J. C., Conner, J. S., & Seborg, D. E. (2007) · Biotechnology Progress · doi:10.1021/bp070063m · Cited by 54 · see citing works
NS0 cell damage by high gas velocity sparging in protein‐free and cholesterol‐free cultures
Zhu, Y., Cuenca, J. V., Zhou, W., & Varma, A. H. (2008) · Biotechnology and Bioengineering · doi:10.1002/bit.21950 · Cited by 53 · see citing works
Thomassen, Y. E., van Sprang, E. N. M., van der Pol, L. A., & Bakker, W. A. M. (2010) · Biotechnology and Bioengineering · doi:10.1002/bit.22788 · Cited by 46 · see citing works
Fault Detection and Diagnosis in an Industrial Fed-Batch Cell Culture Process
Gunther, J. C., Conner, J. S., & Seborg, D. E. (2007) · Biotechnology Progress · doi:10.1002/bp070063m · Cited by 43 · see citing works
Selvarasu, S., Kim, D. Y., Karimi, I. A., & Lee, D. (2010) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2010.07.016 · Cited by 39 · see citing works
Engineering Considerations for Process Development in Mammalian Cell Cultivation
Zhang, H., Wang, W., Quan, C., & Fan, S. (2010) · Current Pharmaceutical Biotechnology · doi:10.2174/138920110790725320 · Cited by 38 · see citing works
Gas holdup in pneumatic reactors
Chisti, Y., & Moo‐Young, M. (1988) · The Chemical Engineering Journal · doi:10.1016/0300-9467(88)80073-x · Cited by 34 · see citing works
An integrated approach to optimization of Escherichia coli fermentations using historical data
Coleman, M. C., Buck, K. K. S., & Block, D. E. (2003) · Biotechnology and Bioengineering · doi:10.1002/bit.10719 · Cited by 31 · see citing works
Advanced methods for bioreactor characterization
Lübbert, A. (1992) · Journal of Biotechnology · doi:10.1016/0168-1656(92)90113-n · Cited by 19 · see citing works
Mahoney, M. J., & Saltzman, W. M. (1999) · Biotechnology and Bioengineering · doi:10.1002/(sici)1097-0290(19990220)62:4 · Cited by 15 · see citing works
Farrell, P., Sun, J., Gao, M., Sun, H., Pattara, B., Zeiser, A., & D’Amore, T. (2012) · Vaccine · doi:10.1016/j.vaccine.2012.05.031 · Cited by 12 · see citing works
Harmsen, M., Stofferis, J., & Malphettes, L. (2011) · BMC Proceedings · doi:10.1186/1753-6561-5-s8-p70 · Cited by 8 · see citing works
Rathore, A. S., & Sofer, G. (2012) · CRC Press eBooks · doi:10.1201/b12013-16 · Cited by 5 · see citing works
Flagged first, then by citations
1 of the 37 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Bioprocessing in the Digital Age: The Role of Process Models
Narayanan, H., Luna, M. F., von Stosch, M., Bournazou, M. N. C., Polotti, G., Morbidelli, M., Butté, A., & Sokolov, M. (2019) · Biotechnology Journal · doi:10.1002/biot.201900172 · Cited by 254 · see citing works
Musa, M., Ayoko, G. A., Ward, A., Rösch, C., Brown, R. J., & Rainey, T. J. (2019) · Cells · doi:10.3390/cells8080851 · Cited by 97 · see citing works
Khuat, T. T., Bassett, R., Otte, E. A., Grevis-James, A., & Gabryś, B. (2024) · Computers & Chemical Engineering · doi:10.1016/j.compchemeng.2024.108585 · Cited by 98 · see citing works
Xu, P., Clark, C., Ryder, T., Sparks, C., Zhou, J., Wang, M., Russell, R., & Scott, C. (2017) · Biotechnology Progress · doi:10.1002/btpr.2417 · Cited by 83 · see citing works
Janakiraman, V., Kwiatkowski, C., Kshirsagar, R., Ryll, T., & Huang, Y. (2015) · Biotechnology Progress · doi:10.1002/btpr.2162 · Cited by 72 · see citing works
Self-citationScale‐Down Model Development in ambr systems: An Industrial Perspective
Sandner, V., Pybus, L. P., McCreath, G., & Glassey, J. (2019) · Biotechnology Journal · doi:10.1002/biot.201700766 · Cited by 63 · see citing works
An Industrial Perspective on Scale-Down Challenges Using Miniaturized Bioreactors
Tajsoleiman, T., Mears, L., Krühne, U., Gernaey, K. V., & Cornelissen, S. (2019) · Trends in biotechnology · doi:10.1016/j.tibtech.2019.01.002 · Cited by 55 · see citing works
Advances in bioreactor control for production of biotherapeutic products
Nikita, S., Mishra, S., Gupta, K., Runkana, V., Gomes, J., & Rathore, A. S. (2023) · Biotechnology and Bioengineering · doi:10.1002/bit.28346 · Cited by 47 · see citing works
Microfluidic tools toward industrial biotechnology
Oliveira, A. F., Pessoa, A. C. S. N., Bastos, R. G., & de la Torre, L. G. (2016) · Biotechnology Progress · doi:10.1002/btpr.2350 · Cited by 45 · see citing works
Digitally enabled approaches for the scale up of mammalian cell bioreactors
Alavijeh, M. K., Baker, I., Lee, Y. Y., & Gras, S. L. (2022) · Digital Chemical Engineering · doi:10.1016/j.dche.2022.100040 · Cited by 44 · see citing works
Novel scale-up strategy based on three-dimensional shear space for animal cell culture
Li, C., Teng, X., Peng, H., Yi, X., Zhuang, Y., Zhang, S., & Xia, J. (2019) · Chemical Engineering Science · doi:10.1016/j.ces.2019.115329 · Cited by 29 · see citing works
Kreye, S., Stahn, R., Nawrath, K., Goralczyk, V., Zoro, B., & Goletz, S. (2019) · Biotechnology Progress · doi:10.1002/btpr.2832 · Cited by 29 · see citing works
Mora, A., Zhang, S., Carson, G. R., Nabiswa, B., Hossler, P., & Yoon, S. (2017) · Biotechnology Progress · doi:10.1002/btpr.2584 · Cited by 27 · see citing works
Segmented linear modeling of CHO fed‐batch culture and its application to large scale production
Yahia, B. B., Gourévitch, B., Malphettes, L., & Heinzle, E. (2016) · Biotechnology and Bioengineering · doi:10.1002/bit.26214 · Cited by 26 · see citing works
Kinetic modeling of Chinese hamster ovary cell culture: factors and principles
Tang, P., Xu, J., Louey, A., Tan, Z., Yongky, A., Liang, S., Li, Z. J., Weng, Y., & Liu, S. (2020) · Critical Reviews in Biotechnology · doi:10.1080/07388551.2019.1711015 · Cited by 26 · see citing works
Ahuja, S., Jain, S., & Ram, K. (2015) · Biotechnology Progress · doi:10.1002/btpr.2134 · Cited by 22 · see citing works
Chaudhary, G., Luo, R., George, M., Tescione, L. D., Khetan, A., & Lin, H. (2020) · Biotechnology and Bioengineering · doi:10.1002/bit.27314 · Cited by 21 · see citing works
Self-citationTikhomirova, T. S., Taraskevich, M. S., & Ponomarenko, O. V. (2018) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-018-9194-z · Cited by 20 · see citing works
Teke, G. M., Gakingo, G. K., & Pott, R. W. M. (2022) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2022.10.017 · Cited by 20 · see citing works
Nie, L., Gao, D., Jiang, H., Gou, J.-X., Li, L., Hu, F., Guo, T., Wang, H., & Qu, H. (2019) · AAPS PharmSciTech · doi:10.1208/s12249-019-1451-7 · Cited by 19 · see citing works
Li, C., Shu, W., Wang, S., Liu, P., Zhuang, Y., Zhang, S., & Xia, J. (2018) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2018.08.005 · Cited by 18 · see citing works
Zhang, X., Moroney, J., Hoshan, L., Jiang, R., & Xu, S. (2019) · Biochemical Engineering Journal · doi:10.1016/j.bej.2019.107307 · Cited by 16 · see citing works
Analysis of Transformed Upstream Bioprocess Data Provides Insights into Biological System Variation
Richelle, A., Lee, B. W., Portela, R. M. C., Raley, J., & von Stosch, M. (2020) · Biotechnology Journal · doi:10.1002/biot.202000113 · Cited by 12 · see citing works
Teng, X., Li, C., Yi, X., & Zhuang, Y. (2021) · Bioresources and Bioprocessing · doi:10.1186/s40643-021-00393-3 · Cited by 11 · see citing works
Production of Protein Therapeutics in the Quality by Design (QbD) Paradigm
Rathore, A. S., & Singh, S. K. (2016) · Topics in medicinal chemistry · doi:10.1007/7355_2015_5004 · Cited by 11 · see citing works
Microbioreactors for Process Development and Cell-Based Screening Studies
Frey, L. J., & Krull, R. (2020) · Advances in biochemical engineering, biotechnology · doi:10.1007/10_2020_130 · Cited by 10 · see citing works
Practical Considerations for the Scale-Up of Chinese Hamster Ovary (CHO) Cell Cultures
Lemire, L., Pham, P. L., Durocher, Y., & Henry, O. (2021) · Cell engineering · doi:10.1007/978-3-030-79871-0_12 · Cited by 11 · see citing works
Yang, P. Y., Hui, C. J., Tien, D. J., Snowden, A. W., Derfus, G. E., & Opel, C. F. (2019) · Journal of Biotechnology · doi:10.1016/j.btecx.2019.100006 · Cited by 10 · see citing works
Predictive modeling for cell culture in commercial manufacturing of biotherapeutics
Panjwani, S., Almazan, A., Hille, R., & Spetsieris, K. (2024) · Biotechnology and Bioengineering · doi:10.1002/bit.28813 · Cited by 8 · see citing works
Yahia, B. B., Piednoir, A., Dahomais, T., Eggermont, S., & Paul, W. (2023) · Biotechnology and Bioengineering · doi:10.1002/bit.28396 · Cited by 7 · see citing works
Kenmoku, H., Kaneko, A., Saito, T., Nemoto, T., Kato, Y., & Ohira, S. (2025) · Engineering in Life Sciences · doi:10.1002/elsc.70054 · Cited by 6 · see citing works
Zhang, W., Ran, Q., Zhao, L., Wan, Y., Ye, Q., & Tan, W. (2024) · Biochemical Engineering Journal · doi:10.1016/j.bej.2024.109309 · Cited by 6 · see citing works
Huang, Y., & Kwiatkowski, C. (2015) · Pharmaceutical Bioprocessing · doi:10.4155/pbp.15.22 · Cited by 6 · see citing works
Reyes, S., Lemire, L., Molina, R., Roy, M., L′Ecuyer‐Coelho, H., Martynova, Y., Cass, B., Voyer, R., Durocher, Y., Henry, O., et al. (2024) · Biotechnology Progress · doi:10.1002/btpr.3467 · Cited by 6 · see citing works
Reyes, S., Pham, P. L., Durocher, Y., & Henry, O. (2024) · Biotechnology Progress · doi:10.1002/btpr.3507 · Cited by 7 · see citing works
Alhattab, M., & Puri, M. (2024) · Algal Research · doi:10.1016/j.algal.2024.103512 · Cited by 4 · see citing works
Dasgupta, A., Thomas, J. A., Anand, A., DeVincentis, B., McCahill, M., Sood, A., Kinross, J., & Rajendran, A. (2025) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2025.06.012 · Cited by 4 · see citing works
Development and Qualification of a Cell Culture Scale‐Down Model
Khattak, S. F., & Pferdeort, V. A. (2019) · Wiley eBooks · doi:10.1002/9783527811410.ch16 · Cited by 4 · see citing works
Robitaille, J., Manceur, A. P., Rodenbrock, A., & Loignon, M. (2024) · Biochemistry · doi:10.5772/intechopen.114000 · Cited by 3 · see citing works
Banton, D., Canova, C. T., Clark, K., Naguib, S., & Schaefer, E. J. (2020) · Biotechnology and Bioengineering · doi:10.1002/bit.27282 · Cited by 3 · see citing works
Mammalian Fed‐batch Cell Culture for Biopharmaceuticals
Yang, W. C. (2019) · doi:10.1002/9783527811410.ch13 · Cited by 2 · see citing works
Dewatering of microalgae from dilute suspensions for biofuels production: A single stage approach
Musa, M. (2020) · Queensland University of Technology · doi:10.5204/thesis.eprints.198026 · Cited by 2 · see citing works
Reconstruction of Industrial Processes for Cultivating CHO Cells in a Scale-Down Bioreactor Model
Морозов, А. Н., Захаров, З. В., Kalenov, S. V., & Авгатович, Х. Р. (2021) · Biotekhnologiya · doi:10.21519/0234-2758-2021-37-4-112-122 · Cited by 1 · see citing works
Gao, J., Hazeltine, L. B., Stroud, N., Liu, N., & Huang, Y. (2024) · Biotechnology Progress · doi:10.1002/btpr.3423
A CFD‐Based Digital Framework for Scaling Optimization of Orbital Rocking Bioreactors
Ryu, S. H., Kim, Y. J., Jeon, J. H., Ji, M. K., Kim, T. H., Kim, K. N., Yang, D., Kim, T., & Hong, J. K. (2025) · Biotechnology Journal · doi:10.1002/biot.70138
Teng, X., Li, C., Yi, X., & Zhuang, Y. (2021) · Figshare · doi:10.6084/m9.figshare.15162300
- Authors
- Tescione, L., Lambropoulos, J., Paranandi, M. R., Makagiansar, H., & Ryll, T.
- Affiliations
- Biogen (United States) (US)
- Biogen Idec, Cell Culture Development 14 Cambridge Center Cambridge Massachusetts 02142
- Corresponding author
- Tescione, Lia
- Venue
- Biotechnology and Bioengineering
- Publisher
- Wiley
- Published
- 2014-09-26
- Volume
- 112
- Issue
- 1
- Pages
- 84-97
- ISSN
- 0006-3592 (ISSN-L)
- 1097-0290
- OpenAlex
- W2140422856
- Cited by
- 46
- References
- 37
- Citations by year
- 2025: 3 · 2024: 8 · 2023: 2 · 2022: 2 · 2021: 4 · 2020: 6 · 2019: 10 · 2018: 3 · 2017: 1 · 2016: 4 · 2015: 3
- FWCI
- 1.38
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biotechnology and Bioengineering (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › Fluid Dynamics and Mixing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Keywords
- oxygen transfer, volumetric mass transfer coefficient, oxygen transfer rate, sparger design, bubble size, CHO cell culture, recombinant protein expression, viable cell density, product quality prediction
- MeSH terms
- Animals, Cricetulus, Cricetinae, Humans, Models, Biological, Oxygen, Recombinant Proteins, Multivariate Analysis, CHO Cells, Cell Culture Techniques, Bioreactors, Principal Component Analysis
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