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Acoustic Cell Filter for High Density Perfusion Culture of Hybridoma Cells

Bio/Technology volume 12, pages 281–284 (1994)Cite this article

Abstract

We have developed a flow–through device which uses high frequency, low energy ultrasonic resonance fields to transiently aggregate hybridoma cells and return them by sedimentation to a perfusion bioreactor. The system retained up to 99 percent of the inflowing viable cells with no measurable effect on viability. Viable cells were selectively retained at up to 3 percent higher efficiency than nonviable cells. A stirred tank bioreactor was operated for 700 hours with acoustic cell recycle. Concentrations greater than 5×107 cells/ml were attained with a 5–fold increase in antibody concentration and a 70–fold increase in volumetric productivity compared with batch culture.

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References

  1. Feder, J., Tolbert, W.R. 1983. The large-scale cultivation of mammalian cells. Scientific Amer. 248: 24–31.

    Article  Google Scholar 

  2. Reuveny, S., Velez, D., Miller, L. and Macmillan, J.D. 1986. Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors. J. Immunol. Methods. 86: 61–69.

    Article  CAS  Google Scholar 

  3. Hansen, H.A., Damgaard, B. and Emborg, C. 1993. Enhanced antibody production associated with altered amino acid metabolism in a hybridoma high-density perfusion culture established by gravity separation. Cytotechnol. 11: 155–166.

    Article  CAS  Google Scholar 

  4. Himmelferb, P., Thayer, P.S. and Martin, H.E. 1969. Spin filter culture: the propagation of mammalian cells in suspension. Science 164: 555–557.

    Article  Google Scholar 

  5. Avgerinos, G.C., Drapeau, D. and Socolow, J.S., Hsiao, K. and Broeze, R.J. 1990. Spin filter system for high density cell culture: production of recombinant urinary type plasminogen activator in CHO cells. Bio/Technology 8: 54–58.

    CAS  PubMed  Google Scholar 

  6. Esclade, L.R.J., Carrel, S. and Peringer, P. 1991. Influence of screen material on the fouling of spin filters. Biotechnol. Bioeng. 38: 159–168.

    Article  CAS  Google Scholar 

  7. Brennan, A.J., Shevitz, J. and Macmillan, J.D. 1987. A perfusion system for antibody production by shear sensitive hybridoma cells in a stirred reactor. Biotechnol. Tech. 1: 169–174.

    Article  CAS  Google Scholar 

  8. Batt, B.C., Davis, R.H. and Kompala, D.S. 1990. Inclined sedimentation for selective retention of viable hybridomas in a continuous suspension bioreactor. Biotechnol. Prog. 6: 458–464.

    Article  CAS  Google Scholar 

  9. Jager, V. 1987. A novel perfusion system for the large-scale cultivation of animal cells based on a continuous flow centrifuge, p. 397–401 In: Approaches to Animal Cell Technology. R. E. Spier and J. B. Griffiths (Eds.). Butterworth, Oxford.

    Google Scholar 

  10. Tokashiki, M., Arai, T., Hamamoto, K. and Ishimaru, K. 1990. High density culture of hybridoma cells using a perfusion culture vessel with external centrifuge. Cytotechnol. 3: 239–244.

    Article  CAS  Google Scholar 

  11. Kilburn, D.G., Clarke, D.J., Coakley, W.T. and Bardsley, D.W. 1989. Enhanced sedimentation of mammalian cells following acoustic aggregation. Biotechnol. Bioeng. 34: 559–562.

    Article  CAS  Google Scholar 

  12. Hager, F. and Benes, E. 1991. A summary of all forces acting on spherical particles in a sound field. Ultrasonics International 91 Conference Proceedings, Le Touquet, France, p. 283–286.

  13. Whitworth, G. and Coakley, W.T. 1992. Particle column formation in a stationary ultrasonic field. J. Acoust. Soc. Am. 91: 79–85.

    Article  Google Scholar 

  14. Rooney, J.A. 1991. Other non-linear acoustic phenomena, p. 65–96 In: Ultrasound: Its Chemical, Physical and Biological Effects. K. S. Suslick (Ed.). VCH Publishers, New York 1988.

    Google Scholar 

  15. Schram, C.J. Manipulation of particles in an acoustic field, p. 293–322 In: Advance in Sonchemistry, Vol. 2. T.J.Mason (Ed).JAI Press Ltd., London.

  16. Mandralis, Z.I., Feke, D.I. and Alder, R.J. 1990. Transient response of fine particle suspensions to mild planar ultrasonic fields. J. Fluid/Particle Separ. 373: 115–121.

    Google Scholar 

  17. Apfel, R.E. 1990. Acoustic radiation pressure—principles and application to separation science. Fortschritte der Akustic DAGA 90 S.19–26

    Google Scholar 

  18. Ziltner, H.J., Clark-Lewis, I., Fasekas, B., Groth, S., Orban, P.C., Hood, L.E., Kent, S.B.H. and Schrader, J.W. 1988. Monoclonalantipeptideantibodies recognize IL-3 and neutralize its bioactivity in vivo. J. Immunol. 4: 1182–1187.

    Google Scholar 

  19. Jervis, E. and Kilburn, D.G. 1991. Rapid immunoassay technique for process monitoring of animal cell fermentations. Biotechnol. Prog. 7: 28–32.

    Article  CAS  Google Scholar 

  20. Nowotny, H. and Benes, E. 1987. General one-dimensional treatment of layered piezoelectric resonators with two electrodes. J. Acoust. Soc. Am. 82: 513–521.

    Article  Google Scholar 

  21. Schmid, M., Benes, E. and Sedlaczek, R. 1990. A computer controlled system for the measurement of complete admittance spectra of piezoelectric resonators. Meas. Sci. Technol. 1: 970–975.

    Article  Google Scholar 

  22. Trampler, F., Benes, E., Burger, W. and Groeschl, M. 1993. Multilayered piezoelectric resonator for the separation of suspended particles. Austrian Patent Appl. No. A926/93.

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Author notes

  1. James M. Piret: Corresponding author.

Authors and Affiliations

  1. Biotechnology Laboratory & Department of Chemical Engineering, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada

    James M. Piret

  2. Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada

    Stefan A. Sonderhoff & Douglas G. Kilburn

  3. Sonofloc Biotech Inc., Richmond, BC, V6V 1Z3, Canada

    Felix Trampler, Stefan A. Sonderhoff & Phylis W. S. Pui

  4. Institute for General Physics, TechnicalUnivesity of Vienna, Vienna, A-1040, Austria

    Felix Trampler

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  1. Felix Trampler
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  2. Stefan A. Sonderhoff
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  3. Phylis W. S. Pui
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  4. Douglas G. Kilburn
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  5. James M. Piret
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Trampler, F., Sonderhoff, S., Pui, P. et al. Acoustic Cell Filter for High Density Perfusion Culture of Hybridoma Cells. Nat Biotechnol 12, 281–284 (1994). https://doi.org/10.1038/nbt0394-281

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