Protein Adsorption and Hydrodynamic Stability of a Dense, Pellicular Adsorbent in High-Biomass Expanded Bed Chromatography

  • Chow, Yen Mei (Department of Process and Food Engineering, Faculty of Engineering, University Putra Malaysia) ;
  • Tey, Beng Ti (Department of Chemical and Environmental Engineering, Faculty of Engineering, University Putra Malaysia) ;
  • Ibrahim, Mohd Nordin (Department of Process and Food Engineering, Faculty of Engineering, University Putra Malaysia) ;
  • Ariff, Arbakariya (Department of Bioprocess Technology, Faculty of Biotechnology and Molecular Science, University Putra Malaysia) ;
  • Ling, Tae Chuan (Department of Process and Food Engineering, Faculty of Engineering, University Putra Malaysia)
  • 발행 : 2006.06.30

초록

A dense, pellicular UpFront adsorbent ($p=1.5 g/cm^3$, UpFront Chromatography, Cophenhagen, Denmark) was characterized in terms of hydrodynamic properties and protein adsorption performance in expanded bed chromatography. Cibacron Blue 3GA was immobilised into the adsorbent and protein adsorption of bovine serum albumin (BSA) was selected to test the setup. The Bodenstein number and axial dispersion coefficient estimated for this dense pellicular adsorbent was 54 and $1.63{\times}10^{-5}m^2/s$, respectively, indicating a stable expanded bed. It could be shown that the BSA protein was captured by the adsorbent in the presence of 30% (w/v) of whole-yeast cells with an estimated dynamic binding capacity $(C/C_o = 0.01)$ of approximately 6.5 mg/mL adsorbent.

키워드

참고문헌

  1. Chow, Y. M., M. N. Ibrahim, B. T. Tey, A. B. Ariff, and T. C. Ling (2005) The influence of bakers' yeast cells on protein adsorption in anion exchange expanded bed chromatography. Biotechnol. Bioprocess Eng. 10: 280-283 https://doi.org/10.1007/BF02932026
  2. Chow, Y. M., M. N. Ibrahim, B. T. Tey, A. B. Ariff, and T. C. Ling (2005) The influence of bakers' yeast cells on protein adsorption in dye-ligand expanded bed chromatography. Biotechnol. Bioprocess Eng. 10: 552-555 https://doi.org/10.1007/BF02932293
  3. Ling, T. C., C. K. Loong, W. S. Tan, B. T. Tey, W. M. W. Abdullah, and A. Ariff (2004) Purification of filamentous bacteriophage M13 by expanded bed anion exchange chromatography. J. Microbiol. 42: 228-232
  4. Tan, Y. P., T. C. Ling, K. Yusoff, W. S. Tan, and B. T. Tey (2005) Comparative evaluation of three purification methods for the nucleocapsid protein of Newcastle disease virus from Escherichia coli homogenates. J. Microbiol. 43: 295-300
  5. Thommes J., M. 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 https://doi.org/10.1002/bit.260450304
  6. Hjorth, R. (1997) Expanded-bed adsorption in industrial bioprocessing: recent developments. Trends Biotechnol. 15: 230-235 https://doi.org/10.1016/S0167-7799(97)01045-7
  7. Ling, T. C. and A. Lyddiatt (2005) Process intensification of fluidized bed dye-ligand adsorption of G3PDH from unclarified disrupted yeast: a case study of the performance of a high-density steel-agarose pellicular adsorbent. Protein Expr. Purif. 42: 160-165 https://doi.org/10.1016/j.pep.2005.03.007
  8. Ling, T. C. (2002) Development of Rapid and Selective Method for the Direct Recovery of Intracellular Protein from Bakers' Yeast. Ph.D. Thesis. The University of Birmingham, Birmingham, UK
  9. 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: A. W. Nienow (eds.). Bioreactor/ Process Fluid Dynamics. BHR Group/Mechanical Engineering Publications, London, UK
  10. Dean, P. D. G. and D. H. Watson (1979) Protein purification using immobilized traizine dyes. J. Chromatogr. 165: 301-319 https://doi.org/10.1016/S0021-9673(00)88187-X
  11. Chambers, G. K. (1977) Determination of Cibacron Blue F3gA substitution in blue Sephadex and blue dextran- Sepharose. Anal. Biochem. 83: 551-556 https://doi.org/10.1016/0003-2697(77)90058-6
  12. Barnfield Frej, A. K., H. J. Johansson, S. Johansson, and P. Leijon (1997) Expanded bed adsorption at production scale: Scale-up verification, process example, and sanitization of column and adsorbent. Bioprocess Biosyst. Eng. 16: 57-63
  13. Amersham Biosciences (1997) Expanded Bed Adsorption: Principles and Methods, Handbooks. pp. 14-16. Uppsala, Sweden
  14. Richardson, J. F. and W. N. Zaki (1954) Sedimentation and fluidization: Part I. Trans. Inst. Chem. Eng. 32: 35-52
  15. Nayak, D. P., S. Ponrathnam, and C. R. Rajan (2001) Macroporous copolymer matrix. IV. Expanded bed adsorption application. J. Chromatogr. A 922: 63-76 https://doi.org/10.1016/S0021-9673(01)00923-2
  16. Draeger, N. M. and H. A. Chase (1991) Liquid fluidized bed adsorption of protein in the presence of cells. Bioseparation 2: 67-80
  17. 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 https://doi.org/10.1002/(SICI)1097-0290(19960305)49:5<512::AID-BIT4>3.0.CO;2-M
  18. Ameskamp, N., C. Priesner, J. Lehmann, and D. Lutkemeyer (1999) Pilot scale recovery of monoclonal antibodies by expanded bed ion exchange adsorption. Bioseparation 8: 169-188 https://doi.org/10.1023/A:1008063626294
  19. Thommes J., M. Weiher, A.Karau, and M. R. Kula (1995) Hydrodynamics and performance in fluidized bed adsorption. Biotechnol. Bioeng. 48: 367-374 https://doi.org/10.1002/bit.260480409
  20. Anspach, F. B., D. Curbelo, R. Hartmann, G. Garke, and W. D. Deckwer (1999) Expanded-bed chromatography in primary protein purification. J. Chromatogr. A 865: 129-44 https://doi.org/10.1016/S0021-9673(99)01119-X
  21. Tong, X. D. and Y. Sun (2002) Particle size and density distributions of two dense matrices in an expanded bed system. J. Chromatogr. A 977: 173-183 https://doi.org/10.1016/S0021-9673(02)01390-0
  22. Bruce, L. J. and H. A. Chase (2001) Hydrodynamics and adsorption behaviour within an expanded bed adsorption column studied using in-bed sampling. Chem. Eng. Sci. 56: 3149-3162 https://doi.org/10.1016/S0009-2509(00)00549-2
  23. Bascoul, A., J. P. Couderc, and H. Delmas (1993) Mouvement des particules solides en fluidisation liquide solide. Chem. Eng. J. 51: 135-150 https://doi.org/10.1016/0300-9467(93)80024-I
  24. Kaufmann, M. (1997) Unstable proteins: how to subject them to chromatographic separations for purification procedures. J. Chromatogr. B 699: 347-369 https://doi.org/10.1016/S0378-4347(96)00512-9