Biotechnology and Bioprocess Engineering

, Volume 6, Issue 6, pp 419–425 | Cite as

Fabrication and characterisation of a novel pellicular adsorbent customised for the effective fluidised bed adsorption of protein products

  • Yan Sun
  • Andrzej W. Pacek
  • Alvin W. Nienow
  • Andrew LyddiattEmail author


A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zirconia-silica (ZS) spheres by water-in-oil emulsification. The agarose evenly laminated the ZS bead to a depth of 30 μm, and the resulting pellicular assembly was characterised by densities up to 2.39 g/mL and a mean particle diameter of 136 μm. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark adsorbents necessary to achieve common values of bed expansion. Furthermore, the pellicular particles were characterised by improved qualities of chromatographic behaviour, particularly with respect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorption were attributed to the physical design and pellicular deployment of the reactive surfaces in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks (whole fermentation broths, cell disruptates and biological extracts).


pellicular adsorbent adsorption equilibrium adsorption kinetics fluidised bed hydrodynamics 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    Wells, C. M., K. Patel, and A. Lyddiatt (1987) Liquid fluidised bed adsorption. pp. 200–209. In: M. J. Verrall and M. J. Hudson (eds.):Separations for Biotechnology: Ellis-Horwood, Chichester, UK.Google Scholar
  2. [2]
    Draeger N. M., and H. A. Chase (1990) Liquid fluidised bed adsorption in the presence of cells.Bioseparation 2: 67–80.Google Scholar
  3. [3]
    Hansson M., S. Stahl, R. Hjorth, M. Uhlen, and T. Mokes (1994) Single-step recovery of a secreted recombinant protein by expanded bed adsorption.Bio/Technology 12: 285–288.CrossRefGoogle Scholar
  4. [4]
    Chang Y. K., G. E. McCreath, and H. A. Chase (1995) Development of an expanded bed technique for an affinity purification of G6PDH from unclarified yeast cell homogenates.Biotechnol. Bioeng. 48: 355–366.CrossRefGoogle Scholar
  5. [5]
    Thoemmes J., S. Halfar, S. Lenz, and M-R. Kula (1995) Purification of monoclonal antibodies from whole hybridoma fermentation broth by fluidized bed adsorption.Biotechnol. Bioeng. 45: 205–211.CrossRefGoogle Scholar
  6. [6]
    Chang Y. K., and H. A. Chase (1996) Ion exchange purification of G6PDH from unclarified yeast cell homogenates using expanded bed adsorption.Biotechnol. Bioeng. 49: 204–216.CrossRefGoogle Scholar
  7. [7]
    Hamilton G. E., P. H. Morton, T. W. Young, and A. Lyddiart (1999) Process intensification by direct product sequestration from batch fermentations: Application of a fluidized bed, multi-bed external loop contactor.Biotechnol. Bioeng. 64: 310–321.CrossRefGoogle Scholar
  8. [8]
    Hamilton, G. E., F. Luechau, S. C. Burton, and A. Lyddiatt (2000) Development of a mixed mode adsorption process for the direct sequestration of an extracellular protease from microbial batch cultures.J. Biotechnol. 79: 103–115.CrossRefGoogle Scholar
  9. [9]
    Bierau, H., R. Hinton, and A. Lyddiatt, A. (2001) Direct process integration of cell disruption and fluidised bed adsorption in the recovery of labile microbial enzymes.Bioseparation 10: 73–85.CrossRefGoogle Scholar
  10. [10]
    Hjorth, R. (1997) Expanded bed adsorption in industrial bioprocessing: Recent developments.Trends Biotechnol. 15: 230–235.CrossRefGoogle Scholar
  11. [11]
    Chase, H. A. (1994) Purification of proteins by adsorption chro-matography in expanded beds.Trends Biotechnol. 12: 296–303.CrossRefGoogle Scholar
  12. [12]
    Voute, N., and E. Boschetti (1999) Highly dense beaded sorbents suitable for fluidised bed applications.Bioseparation 8: 115–120.CrossRefGoogle Scholar
  13. [13]
    Gilchrist G. R., M. T. Burns, and A. Lyddiatt (1994) Solid phases for protein adsorption in fluidised beds: Comparison of commercial and custom assembled particles. pp. 186–192. In: M. J. Verrall and M. J. Hudson (eds.),Separations for Biotechnology: Ellis-Horwood, Chichester, UK.Google Scholar
  14. [14]
    Finette G. M. S., Q-M. Mao, and M. T. W. Hearn (1998) Examination of protein adsorption in fluidized bed and packed bed columns at different temperature using frontal chromatographic methods.Biotechnol. Bioeng. 58: 35–46.CrossRefGoogle Scholar
  15. [15]
    Thoemmes J., M. Weiher, A. Karau, and M-R. Kula (1995) Hydrodynamics and performance in fluidized bed adsorption.Biotechnol. Bioeng. 48: 367–374.CrossRefGoogle Scholar
  16. [16]
    McCreath G. E., H. A. Chase, R. O. Owen, and C. R. Lowe (1995) Expanded bed affinity chromatography of dehydrogenase from baker's yeast using dye-ligand perfluoropolymer supports.Biotechnol. Bioeng. 48: 341–354.CrossRefGoogle Scholar
  17. [17]
    Zhu J., A. Lyddiatt, A. W. Pacek, and A. W. Nienow (1997) Fabrication and characterisation of agar/zircon sand composite adsorbents for protein recovery in liquid fluidised beds. pp 103–114. In: Proceedings Fourth International Conference on Bioreactor and Bioprocess Fluid Dynamics. BHR Group/Inst. Mechanical Engineering Publications, London, UK.Google Scholar
  18. [18]
    Gibson, N. B. (1991).Liquid Fluidised Bed Adsorption in Protein Recovery: Assembly and Characterisation of Dedicated Solid Phases. PhD thesis, University of Birmingham, UK.Google Scholar
  19. [19]
    Gibson, N. B., and A. Lyddiatt (1993) Cellulose composites in liquid fluidised bed adsorption and recovery of proteins. pp. 55–62. In: J. F. Kennedy, G. O. Phillips and P. A. Williams (eds.).Cellulosics Materials for Selective Separations and Other Technologies, Ellis-Horwood, Chichester, UKGoogle Scholar
  20. [20]
    Lyddiatt A., and D. A. O'Sullivan DA (1998) Biochemical recovery and purification of gene therapy vectors.Curr. Opin. Biotechnol. 9: 177–185.CrossRefGoogle Scholar
  21. [21]
    He, L-Z., Y-R. Gan, and Y. Sun (1997) Adsorption-desorption of BSA to highly-substituted dye-ligand adsorbents: quantitative study of the effect of ionic strength.Bioprocess Eng. 17: 301–305.CrossRefGoogle Scholar
  22. [22]
    Wright, P. R., F. J. Muzzio, and B. J. Glasser (1993) Batch uptake of lysozyme: Effect of solution viscosity and mass transfer on adsorption.Biotechnol. Prog. 14: 913–921.CrossRefGoogle Scholar
  23. [23]
    Levenspiel, O. (1999)Chemical Reaction Engineering, 3rd ed, John Wiley & Sons, New York, USA.Google Scholar
  24. [24]
    Barnfield Frej, A-K., H. J. Johansson, S. Johansson, S, and P. Leijon (1997) Expanded bed adsorption at production scale: Scale up verification, process example and sanitisation of column and adsorbent.Bioprocess Eng. 15: 57–61.CrossRefGoogle Scholar
  25. [25]
    Sexena, V. P., and D. B. Wetlaufer (1970) Formation of three dimensional structure in proteins: I, rapid nonenzymic reactivation of reduced lysozyme.Biochemistry 9: 5015–5022.CrossRefGoogle Scholar
  26. [26]
    Bird, B., W. E. Stewart, and I. Lighfoot (1960)Transport Phenomena, Wiley and Sons, NY, USA.Google Scholar
  27. [27]
    Geankopolis C. J. (1983)Transport Processes and Unit Operations. 2nd ed. Allyn and Bacon, NY, USA.Google Scholar
  28. [28]
    Tyn M. T. and T. W. Guesk (1990) Prediction of diffusion coefficients of proteins.Biotechnol. Bioeng. 35: 326–338.CrossRefGoogle Scholar
  29. [29]
    Clemmitt, R. H., L. J. Bruce, and H. A. Chase (1999) Online monitoring of the purification of GST-(His)6 from an unclarifiedEscherichia coli homogenate within an immobilised metal affinity expanded bed.Bioseparation 8: 53–67.CrossRefGoogle Scholar
  30. [30]
    Richardson, J. F. and W. N. Zaki (1954) Sedimentation and fluidization: Part I.Trans. Inst. Chem. Eng. 32: 35–53.Google Scholar
  31. [31]
    Chang, Y. K., and H. A. Chase (1996) Development of operating conditions for protein purification using expanded bed techniques: the effect of the degree of bed expansion on adsorption performance.Biotechnol. Bioeng. 49: 512–526.CrossRefGoogle Scholar
  32. [32]
    Bierau, H., Z. Zhang, and A. Lyddiatt (1999) Direct process integration of cell disruption and fluidised bed adsorption for the recovery of intracellular proteins.J. Chem. Technol. Biotechnol. 74: 208–212.CrossRefGoogle Scholar

Copyright information

© The Korean Society for Biotechnology and Bioengineering 2001

Authors and Affiliations

  • Yan Sun
    • 2
  • Andrzej W. Pacek
    • 1
  • Alvin W. Nienow
    • 1
  • Andrew Lyddiatt
    • 1
    Email author
  1. 1.Biochemical Recovery Group, Centre for Formulation Engineering, School of Engineering (Chemical Engineering)University of BirminghamBirminghamUK
  2. 2.Department of Biochemical EngineeringTianjin UniversityTianjinP. R. China

Personalised recommendations