Skip to main content
SpringerLink
Log in

On-line monitoring of infected Sf-9 insect cell cultures by scanning permittivity measurements and comparison with off-line biovolume measurements

  • Special Issue JAACT
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Two infected Sf-9 cell cultures were monitored on-line by multi-frequency permittivity measurements using the Fogale BIOMASS SYSTEM® and by applying different off-line methods (CASY®1, Vi-CELL™, packed cell volume) to measure the biovolume and the mean diameter of the cell population. During the growth phase and the early infection phase the measured permittivity at the working frequency correlated well with the different off-line methods for the biovolume. We found a value of 0.67 pF cm−1 permittivity per unit of total biovolume (CASY) (μL mL−1). After the maximum value in the permittivity was reached, i.e. when the viability of the cultures decreased significantly, we observed different time courses for the biovolume depending on the applied method. The differences were compared and could be explained by the underlying measurement principles. Furthermore, the characteristic frequency (fC) was calculated from the on-line scanning permittivity measurements. The fC may provide an indication of changes in cell diameter and membrane properties especially after infection and could also be an indicator for the onset of the virus production phase. The changes in fC were qualitatively explained by the underlying equation that is correlating fC and the properties of the cell population (cell diameter, intracellular conductivity and capacitance per membrane area).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

α:

Cole–Cole alpha

CM :

Capacitance per membrane area (F m−2)

Δε:

Cell permittivity (pF cm−1)

ΔεFOGALE :

Permittivity difference at f1 and f2 (pF cm−1)

fC :

Characteristic frequency (Hz)

hema:

Hemacytometer

KLa:

Volumetric oxygen mass transfer coefficient (h−1)

PBS:

Phosphate buffered saline

PCV:

Packed cell volume (μL mL−1)

TOI:

Time of infection with baculovirus (h)

References

  • Ansorge S, Esteban G, Ghommidh C, Schmid G (2007) Monitoring nutrient limitations by online capactitance measurements in batch and fed-batch CHO fermentations. In: Smith R (ed) Conference Proceedings to the 19th ESACT Meeting: Cell Technology for Cell Products. Springer, Dordrecht/NL, pp 723–726

  • Cannizzaro C, Gugerli R, Marison I, von Stockar U (2003) On-line biomass monitoring of CHO perfusion culture with scanning dielectric spectroscopy. Biotechnol Bioeng 84:597–610

    Article  CAS  Google Scholar 

  • Chico E, Jäger V (1998) Measurements of changes in cell size distribution to monitor Baculovirus infection of insect cells. In: Merten OW, Perrin P, Griffiths B (eds) New developments and new applications in animal cell technology. Kluwer Academic Publishers, Dordrecht, 329–331

    Google Scholar 

  • Cook JA, Mitchell JB (1989) Viability measurements in mammalian cell systems. Anal Biochem 179:1–7

    Article  CAS  Google Scholar 

  • Davey CL (1993) The biomass monitor source book. Department of Biological Sciences, University of Wales, Aberystwyth

    Google Scholar 

  • Davey CL, Marckx GH, Kell DB (1993) On the dielectric method of monitoring cellular viability. Pure Appl Chem 65:1921–1926

    Article  CAS  Google Scholar 

  • Ducommun P, Bolzonella I, Rhiel M, Pugeaud P, von Stockar U, Marison IW (2001) On-line determination of animal cell concentration. Biotechnol Bioeng 72:515–522

    Article  CAS  Google Scholar 

  • Ducommun P, Kadouri A, von Stockar U, Marison IW (2002) On-line determination of animal cell concentration in two industrial high-density culture processes by dielectric spectroscopy. Biotechnol Bioeng 77:316–323

    Article  CAS  Google Scholar 

  • Fehrenbach R, Comberbach M, Pêtre JO (1992) On-line biomass monitoring by capacitance measurement. J Biotechnol 23:303–314

    Article  CAS  Google Scholar 

  • Fogale Nanotech (2004) Biomass System User Manual V 3.0

  • Foster KR, Schwan HP (1989) Dielectric properties of tissues and biological materials: A critical review. Crit Rev Biomed Eng 17:25–104

    CAS  Google Scholar 

  • Guan Y, Evans PM, Kemp RB (1998) Specific heat flow rate: an on-line monitor and potential control variable of specific metabolic rate in animal cell culture that combines microcalorimetry with dielectric spectroscopy. Biotechnol Bioeng 58:464–477

    Article  CAS  Google Scholar 

  • Harris CM, Todd RW, Bungard SJ, Lovitt RW, Morris JG, Kell DB (1987) Dielectric permittivity of microbial suspensions at radio frequencies: a novel method for the real-time estimation of microbial biomass. Enzyme Microb Technol 9:181–186

    Article  CAS  Google Scholar 

  • Kamen AA, Bédard C, Tom R, Perret S, Jardin B (1996) On-line monitoring of respiration in recombinant-baculovirus infected and uninfected insect cell bioreactor cultures. Biotechnol Bioeng 50:36–48

    Article  CAS  Google Scholar 

  • Kost TA, Condreay JP, Jarvis DL (2005) Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 23:567–575

    Article  CAS  Google Scholar 

  • Markx GH, Davey CL (1999) The dielectric properties of biological cells at radiofrequencies: applications in biotechnology. Enzyme Microb Technol 25:161–171

    Article  CAS  Google Scholar 

  • Noll T, Biselli M (1998) Dielectric spectroscopy in the cultivation of suspended and immobilized hybridoma cells. J Biotechnol 63:187–198

    Article  CAS  Google Scholar 

  • Olsson L, Nielsen J (1997) On-line and in situ monitoring of biomass in submerged cultivations. Trends Biotechnol 15:517–522

    Article  CAS  Google Scholar 

  • Ooi BG, Miller LK (1988) Regulation of host RNA levels during baculovirus infection. Virology 166:515–523

    Article  CAS  Google Scholar 

  • Pethig R, Kell DB (1987) The passive electrical properties of biological systems: their significance in physiology, biophysics and biotechnology. Phys Med Biol 32:933–970

    Article  CAS  Google Scholar 

  • Schlaeger E-J (1996) Medium design for insect cell culture. Cytotechnology 20:57–70

    Article  CAS  Google Scholar 

  • Schmid G (1996) Insect cell cultivation: growth and kinetics. Cytotechnology 20:43–56

    Article  CAS  Google Scholar 

  • Schmid G, Wild N, Fountoulakis M, Gallati H, Gentz R, Ozmen L, Garotta G (1994) Production and characterization of soluble mouse and human interferon-g receptors (IFNg-R) from insect cells. In: Spier RE, Griffiths JB, Berthold W (eds) Conference Proceedings to the 12th ESACT Meeting: Animal cell technology: products for today, prospects for tomorrow. Butterworth-Heinemann, Oxford/UK, pp 625–632

  • Schopf B, Howaldt MW, Bailey JE (1990) DNA distribution and respiratory activity of Spodoptera frugiperda populations infected with wild-type and recombinant Autographa californica nuclear polyhedrosis virus. J Biotechnol 15:169–186

    Article  CAS  Google Scholar 

  • Sonnleitner B, Locher G, Fiechter A (1992) Biomass determination. J Biotechnol 25:5–22

    Article  CAS  Google Scholar 

  • Stettler M, Jaccard N, Hacker D, De Jesus M, Wurm FM, Jordan M (2006) New disposable tubes for rapid and precise biomass assessment for suspension cultures of mammalian cells. Biotechnol Bioeng 95:1228–1233

    Article  CAS  Google Scholar 

  • Taticek RA, Shuler ML (1997) Effect of elevated oxygen and glutamine levels on foreign protein production at high cell densities using the insect cell-baculovirus expression system. Biotechnol Bioeng 54:142–152

    Article  CAS  Google Scholar 

  • Winkelmeier P, Glauner B, Lindl T (1993) Quantification of cytotoxicity by cell volume and cell proliferation. Alterna Lab Anim 21:269–280

    Google Scholar 

  • Wong TK, Nielsen LK, Greenfield PF, Reid S (1994) Relationship between oxygen uptake rate and time of infection of Sf9 insect cells infected with a recombinant baculovirus. Cytotechnology 15:157–167

    Article  CAS  Google Scholar 

  • Zeiser A, Bédard C, Voyer R, Jardin B, Tom R, Kamen AA (1999) On-line monitoring of the progress of infection in Sf-9 insect cell cultures using relative permittivity measurements. Biotechnol Bioeng 63:122–126

    Article  CAS  Google Scholar 

  • Zeiser A, Elias CB, Voyer R, Jardin B, Kamen AA (2000) On-line monitoring of physiological parameters of insect cell cultures during the growth and infection process. Biotechnol Prog 16:803–808

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to acknowledge C. Ghommidh from Université Montpellier II and M. Biselli from the Aachen University of Applied Sciences (Department Juelich) for the stimulating and fruitful discussions and remarks, M. Foggetta, M. Siegrist, J.-M. Vonach and J.-C. von Bueren for the support with small scale experiments and the setup of the bioreactor system and H. Remy for the use and help with the CASY®1 system. S. Ansorge was supported from March to September 2004 by a Hoffmann-La Roche AG fellowship and the presented results form a part of his diploma thesis at the Aachen University of Applied Sciences.

Author information

Author notes

  1. Sven Ansorge

    Present address: Animal Cell Technology Group, Biotechnology Research Institute, National Research Council Canada, 6100 Royalmount Avenue, Montreal, QC, Canada, H4P 2R2

Authors and Affiliations

  1. Pharmaceuticals Division, F. Hoffmann-La Roche AG, Pharma Research Basel, Protein Sciences, Basel, 4070, Switzerland

    Sven Ansorge & Georg Schmid

  2. FOGALE Nanotech, Nimes, France

    Geoffrey Esteban

Authors
  1. Sven Ansorge
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Geoffrey Esteban
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Georg Schmid
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sven Ansorge.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ansorge, S., Esteban, G. & Schmid, G. On-line monitoring of infected Sf-9 insect cell cultures by scanning permittivity measurements and comparison with off-line biovolume measurements. Cytotechnology 55, 115–124 (2007). https://doi.org/10.1007/s10616-007-9093-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-007-9093-0

Keywords

Search

Navigation