The Performance of Anion Exchange Expanded Bed Adsorption Chromatography on the Recovery of G6PDH from Unclarified Feedstock with High Biomass Concentration

  • Chow, Yen Mei (Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia) ;
  • Tey, Beng Ti (Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia) ;
  • Ibrahim, Mohd Nordin (Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia) ;
  • Ariff, Arbakariya (Department of Bioprocess Technology, Faculty of Biotechnology and Molecular Science, Universiti Putra Malaysia) ;
  • Ling, Tau Chuan (Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia)
  • Published : 2006.10.30

Abstract

The bed stability of Streamline DEAE (p = 1.2 g/mL) in a 20mm (i.d.) glass expanded bed contactor, and its performance on the recovery of glucose 6-phosphate dehydrogenase (G6PDH) from unclarified yeast homogenate were investigated. A residence time distribution study showed that a stable expanded bed was achieved. The theoretical plate and Bodenstein numbers determined were 25 and 53, respectively. A recovery yield of 87% and purification factor of 4.1 were achieved in the operation using 5% (w/v) biomass concentration feedstock. The performance of the anion exchange EBAC was still considerable good at a biomass concentration as high as 15% (w/v).

Keywords

References

  1. Bonnerjea, J., S. Oh, M. Hoare, and P. Dunnill (1986) Protein purification: The right step at the right time. Bio/Technol. 4: 954-958 https://doi.org/10.1038/nbt1186-954
  2. 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
  3. Tan, Y. P, T. C. Ling, W. S. Tan, K. Yusoff, and B. T. Tey (2005) Purification of recombinant nucleocapsid protein of Newcastle disease virus from unclarified feedstock using expanded bed adsorption chromatography. Protein Expr. Purif. 46: 114-121 https://doi.org/10.1016/j.pep.2005.06.015
  4. Thommes, J., M. Halfar, S. Lenz, and M.-R. Kula (1995) Purification of monoclonal antibodies from whole hybri-doma fermentation broth by fluidized bed adsorption. Biotechnol. Bioeng. 45: 205-211 https://doi.org/10.1002/bit.260450304
  5. Chow, Y. M., B. T. Tey, M. N. Ibrahim, A. Ariff, and T. C. Ling (2006) Protein adsorption and hydrodynamic stability of a dense, pellicular adsorbent in high-biomass expanded bed chromatography. Biotechnol. Bioprocess Eng. 11: 268-272 https://doi.org/10.1007/BF02932042
  6. Boschetti, E., L. Guerrier, P. Girot, and J. Horvath (1995) Preparative high-performance liquid chromatographic separation of proteins with HyperD ion-exchange supports. J. Chromatogr. B, Biomed. Appl. 664: 225-231 https://doi.org/10.1016/0378-4347(94)00406-U
  7. Chow, Y. M., B. T. Tey, M. N. Ibrahim, A. Ariff, and T C. Ling (2005) The disruption of Saccharomyces cerevisiae cells and release of glucose 6-phosphate dehydrogenase (G6PDH) in a horizontal dyno bead mill operated in continuous recycling mode. Biotechnol. Bioprocess Eng. 10: 284-288 https://doi.org/10.1007/BF02932027
  8. 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 sanitiza-tion of column and adsorbent. Bioprocess Biosyst. Eng. 16: 57-63
  9. 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
  10. Ameskamp, N., C. Priesner, J. Lehmann, and D. Lutke-meyer (1999) Pilot scale recovery of monoclonal antibodies by expanded bed ion exchange adsorption. Bioseparation 8: 169-188 https://doi.org/10.1023/A:1008063626294
  11. 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
  12. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72: 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  13. Thommes, J., A. Bader, M. Halfar, A. Karau, M. R. Kula (1996) Isolation of monoclonal antibodies from cell containing hybridoma broth using a protein A coated adsorbent in expanded beds. J. Chromatogr. A 752: 111-122 https://doi.org/10.1016/S0021-9673(96)00504-3
  14. 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
  15. Chow, Y. M., B. T. Tey, M. N. Ibrahim, A. 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
  16. Chow, Y. M., B. T. Tey, M. N. Ibrahim, A. Ariff, and T. C. Ling (2005) The influence of bakers' yeast cells on protein adsorption performance in dye-ligand expanded bed chromatography. Biotechnol. Bioprocess Eng. 10: 552-555 https://doi.org/10.1007/BF02932293
  17. Fernandez-Lahore, H. M., R. Kleef, M. R. Kula, and J. Thommes (1999) The influence of complex biological feedstock on the fluidization and bed stability in expanded bed adsorption. Biotechnol. Bioeng. 64: 484-496 https://doi.org/10.1002/(SICI)1097-0290(19990820)64:4<484::AID-BIT11>3.0.CO;2-Z
  18. Lu, M.-H., D.-Q. Lin, Y.-C. Wu, J.-X. Yun, L.-H. Mei, and S.-J. Yao (2005) Separation of Nattokinase from Bacillus subtilis fermentation broth by expanded bed adsorption with mixed-mode adsorbent. Biotechnol. Bioprocess Eng. 10: 128-135 https://doi.org/10.1007/BF02932582
  19. Viloria-Cols, M. E., R. Hatti-Kaul, and B. Mattiasson (2004) Agarose-coated anion exchanger prevents cell-adsorbent interactions. J. Chromatogr. A 1043: 195-200 https://doi.org/10.1016/j.chroma.2004.05.086