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Application of dielectric spectroscopy for monitoring high cell density in monoclonal antibody producing CHO cell cultivations

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Abstract

The application of dielectric spectroscopy was frequently investigated as an on-line cell culture monitoring tool; however, it still requires supportive data and experience in order to become a robust technique. In this study, dielectric spectroscopy was used to predict viable cell density (VCD) at industrially relevant high levels in concentrated fed-batch culture of Chinese hamster ovary cells producing a monoclonal antibody for pharmaceutical purposes. For on-line dielectric spectroscopy measurements, capacitance was scanned within a wide range of frequency values (100–19,490 kHz) in six parallel cell cultivation batches. Prior to detailed mathematical analysis of the collected data, principal component analysis (PCA) was applied to compare dielectric behavior of the cultivations. PCA analysis resulted in detecting measurement disturbances. By using the measured spectroscopic data, partial least squares regression (PLS), Cole–Cole, and linear modeling were applied and compared in order to predict VCD. The Cole–Cole and the PLS model provided reliable prediction over the entire cultivation including both the early and decline phases of cell growth, while the linear model failed to estimate VCD in the later, declining cultivation phase. In regards to the measurement error sensitivity, remarkable differences were shown among PLS, Cole–Cole, and linear modeling. VCD prediction accuracy could be improved in the runs with measurement disturbances by first derivative pre-treatment in PLS and by parameter optimization of the Cole–Cole modeling.

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References

  1. Rathore AS, Bhambure R, Ghare V (2010) Process analytical technology (PAT) for biopharmaceutical products. Anal Bioanal Chem 398(1):137–154

    Article  CAS  Google Scholar 

  2. Junker BH et al (1994) On-line and in situ monitoring technology for cell density measurement in microbial and animal cell cultures. Bioprocess Eng 10(5–6):195–207

    Google Scholar 

  3. Carvell J, Dowd J (2006) On-line measurements and control of viable cell density in cell culture manufacturing processes using radio-frequency impedance. Cytotechnology 50(1–3):35–48

    Article  CAS  Google Scholar 

  4. Maskow T et al (2008) Observation of non-linear biomass–capacitance correlations: reasons and implications for bioprocess control. Biosens Bioelectron 24(1):123–128

    Article  CAS  Google Scholar 

  5. Cannizzaro C et al (2003) On-line biomass monitoring of CHO perfusion culture with scanning dielectric spectroscopy. Biotechnol Bioeng 84(5):597–610

    Article  CAS  Google Scholar 

  6. Ducommun P et al (2002) On-line determination of animal cell concentration in two industrial high-density culture processes by dielectric spectroscopy. Biotechnol Bioeng 77(3):316–323

    Article  CAS  Google Scholar 

  7. Ansorge S, Esteban G, Schmid G (2010) Multifrequency permittivity measurements enable on-line monitoring of changes in intracellular conductivity due to nutrient limitations during batch cultivations of CHO cells. Biotechnol Prog 26(1):272–283

    CAS  Google Scholar 

  8. Davey CL et al (1993) Introduction to the dielectric estimation of cellular biomass in real time, with special emphasis on measurements at high volume fractions. Anal Chim Acta 279(1):155–161

    Article  Google Scholar 

  9. Ansorge S et al (2010) Monitoring the cell size distribution of mammalian cell cultures using on-line capacitance measurements. In: Noll T (ed) Cells and culture. Springer, Netherlands, pp 853–859

    Chapter  Google Scholar 

  10. Dabros M et al (2009) Cole–Cole, linear and multivariate modeling of capacitance data for on-line monitoring of biomass. Bioprocess Biosyst Eng 32(2):161–173

    Article  CAS  Google Scholar 

  11. Opel CF, Li J, Amanullah A (2010) Quantitative modeling of viable cell density, cell size, intracellular conductivity, and membrane capacitance in batch and fed-batch CHO processes using dielectric spectroscopy. Biotechnol Prog 26(4):1187–1199

    CAS  Google Scholar 

  12. Birch JR, Racher AJ (2006) Antibody production. Adv Drug Deliv Rev 58(5–6):671–685

    Article  CAS  Google Scholar 

  13. Bleckwenn NA et al (2005) Production of recombinant proteins by vaccinia virus in a microcarrier based mammalian cell perfusion bioreactor. Biotechnol Bioeng 90(6):663–674

    Article  CAS  Google Scholar 

  14. Clincke M-F et al (2011) Study of a recombinant CHO cell line producing a monoclonal antibody by ATF or TFF external filter perfusion in a WAVE BioreactorTM. BMC Proc 5(Suppl 8):P105

    Article  Google Scholar 

  15. Chotteau V, Tördahl K, Perroud P (2009) Study of a perfusion process of Chinese hamster ovary cells by ATF filtration in bioreactor. In: ESACT conference, Dublin, 7–10 June 2009

  16. (2012) Modelling and regularity of nonlinear impulsive switching dynamical system in fed-batch culture. Abstr Appl Anal

  17. Ye J et al (2011) Modelling and well-posedness of a nonlinear hybrid system in fed-batch production of 1,3-propanediol with open loop glycerol input and pH logic control. Nonlinear Anal Real World Appl 12(1):364–376

    Article  CAS  Google Scholar 

  18. Bonham-Carter J et al (2011) The use of the ATF system to culture Chinese hamster ovary cells in a concentrated fed-batch system. BioPharm Int 24(6):42–48

    CAS  Google Scholar 

  19. Ansorge S, Esteban G, Schmid G (2010) On-line monitoring of responses to nutrient feed additions by multi-frequency permittivity measurements in fed-batch cultivations of CHO cells. Cytotechnology 62(2):121–132

    Article  CAS  Google Scholar 

  20. Sauer PW et al (2000) A high-yielding, generic fed-batch cell culture process for production of recombinant antibodies. Biotechnol Bioeng 67(5):585–597

    Article  CAS  Google Scholar 

  21. Seth G et al (2013) Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns. Biotechnol Bioeng 110(5):1376–1385

    Article  CAS  Google Scholar 

  22. Warikoo V et al (2012) Integrated continuous production of recombinant therapeutic proteins. Biotechnol Bioeng 109(12):3018–3029

    Article  CAS  Google Scholar 

  23. Chee Furng Wong D et al (2005) Impact of dynamic online fed-batch strategies on metabolism, productivity and N-glycosylation quality in CHO cell cultures. Biotechnol Bioeng 89(2):164–177

    Article  Google Scholar 

  24. Hossler P, Khattak SF, Li ZJ (2009) Optimal and consistent protein glycosylation in mammalian cell culture. Glycobiology 19(9):936–949

    Article  CAS  Google Scholar 

  25. Borys MC, Linzer DIH, Papoutsakis ET (1993) Culture pH affects expression rates and glycosylation of recombinant mouse placental lactogen proteins by Chinese hamster ovary (CHO) Cells. Nat Biotechol 11(6):720–724

    Article  CAS  Google Scholar 

  26. Hakemeyer C et al (2012) At-line NIR spectroscopy as effective PAT monitoring technique in Mab cultivations during process development and manufacturing. Talanta 90:12–21

    Article  CAS  Google Scholar 

  27. Henriques J et al (2010) Monitoring mammalian cell cultivations for monoclonal antibody production using near-infrared spectroscopy. In: Rao G (ed) Optical sensor systems in biotechnology. Springer, Berlin, pp 29–72

    Google Scholar 

  28. Rodrigues LO et al (2008) The use of NIR as a multi-parametric in situ monitoring technique in filamentous fermentation systems. Talanta 75(5):1356–1361

    Article  CAS  Google Scholar 

  29. Reich G (2005) Near-infrared spectroscopy and imaging: basic principles and pharmaceutical applications. Adv Drug Deliv Rev 57(8):1109–1143

    Article  CAS  Google Scholar 

  30. Saerens L et al (2012) In-line NIR spectroscopy for the understanding of polymer–drug interaction during pharmaceutical hot-melt extrusion. Eur J Pharm Biopharm 81(1):230–237

    Article  CAS  Google Scholar 

  31. Zang H et al (2013) Application of near-infrared spectroscopy combined with multivariate analysis in monitoring of crude heparin purification process. Spectrochim Acta Part A Mol Biomol Spectrosc 109:8–13

    Article  CAS  Google Scholar 

  32. Oshita S et al (2011) Monitoring of ATP and viable cells on meat surface by UV–Vis reflectance spectrum analysis. J Food Eng 107(2):262–267

    Article  CAS  Google Scholar 

  33. Patel P, Markx GH (2008) Dielectric measurement of cell death. Enzym Microb Technol 43(7):463–470

    Article  CAS  Google Scholar 

  34. Davey CL, Kell DB (1998) The influence of electrode polarisation on dielectric spectra, with special reference to capacitive biomass measurements: I. Quantifying the effects on electrode polarisation of factors likely to occur during fermentations. Bioelectrochem Bioenergy 46(1):91–103

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biotechnology, Gedeon Richter Plc, 19-21, Gyömrői út, Budapest, 1103, Hungary

    László Párta, Dénes Zalai, Sándor Borbély & Ákos Putics

  2. Department of Applied Biotechnology and Food Science, Budapest University of Technology and Economics, 4, Szent Gellért tér, Budapest, 1111, Hungary

    László Párta

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  1. László Párta
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  2. Dénes Zalai
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  3. Sándor Borbély
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  4. Ákos Putics
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Correspondence to László Párta.

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Párta, L., Zalai, D., Borbély, S. et al. Application of dielectric spectroscopy for monitoring high cell density in monoclonal antibody producing CHO cell cultivations. Bioprocess Biosyst Eng 37, 311–323 (2014). https://doi.org/10.1007/s00449-013-0998-z

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  • DOI: https://doi.org/10.1007/s00449-013-0998-z

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