Journal of Microbiology

  • 제43권3호
  • /
  • Pages.295-300
  • /
  • 2005
  • /
  • 1225-8873(pISSN)
  • /
  • 1976-3794(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Comparative Evaluation of Three Purification Methods for the Nucleocapsid Protein of Newcastle Disease Virus from Escherichia coli Homogenates

  • Tan Yan Peng (Department of Chemical and Environmental Engineering, Institute of Bioscience, University Putra Malaysia) ;
  • Ling Tau Chuan (Department of Process & Food Engineering, Faculty of Engineering, Institute of Bioscience, University Putra Malaysia) ;
  • Yusoff Khatijah (Institute of Bioscience, Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia) ;
  • Tan Wen Siang (Institute of Bioscience, Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia) ;
  • Tey Beng Ti (Department of Chemical and Environmental Engineering, Institute of Bioscience, University Putra Malaysia)
  • 발행 : 2005.06.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

In the present study, the performances of conventional purification methods, packed bed adsorption (PBA), and expanded bed adsorption (EBA) for the purification of the nucleocapsid protein (NP) of Newcastle disease virus (NDV) from Escherichia coli homogenates were evaluated. The conventional methods for the recovery of NP proteins involved multiple steps, such as centrifugation, precipitation, dialysis, and sucrose gradient ultracentrifugation. For the PBA, clarified feedstock was used for column loading, while in EBA, unclarified feedstock was used. Streamline chelating immobilized with $Ni^{2+}$ ion was used as an affinity ligand for both PBA and EBA. The final protein yield obtained in conventional and PBA methods was $1.26\%$ and $5.56\%$, respectively. It was demonstrated that EBA achieved the highest final protein yield of $9.6\%$ with a purification factor of 7. Additionally, the total processing time of the EBA process has been shortened by 8 times compared to that of the conventional method.

키워드

참고문헌

  1. Ameskamp, N., C. Priesner, J. Lehmann, and D. Lutkemeyer. 1999. Pilot scale recovery of monoclonal antibodies by expanded bed ion exchange adsorption. Bioseparation 8, 169-88 https://doi.org/10.1023/A:1008063626294
  2. Anspach, F.B., D. Curbelo, R. Hartmann, G. Garke, and W.D. Deekwer. 1999. Expanded-bed chromatography in primary protein purification. J. Chromatogr. A 865, 129-144 https://doi.org/10.1016/S0021-9673(99)01119-X
  3. Batt, B.C., V.M. Yabannavar, and V. Singh. 1995. Expanded bed adsorption process for protein recovery from whole mammalian cell culture broth. Bioseparation 5, 41-52
  4. 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
  5. Chaga, G.S. 2001. Twenty-five years of immobilized metal ion affinity chromatography: past, present and future. J. Biochem. Biophys. Methods 49, 313-334 https://doi.org/10.1016/S0165-022X(01)00206-8
  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 https://doi.org/10.1002/(SICI)1097-0290(19960120)49:2<204::AID-BIT9>3.0.CO;2-U
  7. Chase, H.A. and N.M. Draeger. 1992. Affinity purification of proteins using expanded beds. J. Chromatogr. A 597,129-145 https://doi.org/10.1016/0021-9673(92)80103-2
  8. Ersson, B., L. Ryden, and J.-C. Janson. 1998. Introduction to protein purification, p. 3 - 40. In J-C. Janson, L. Ryden (eds.), Protein Purification: Principles, High-Resolution Methods, and Applications. John Wiley & Sons, New York
  9. Hu, Y.C., C. T. Tsai, Y.C. Chung, J.T. Lu, and J.T. Hsu. 2003. Generation of chimeric baculovirus with histidine-tags displayed on the envelope and its purification using immobilized metal affinity chromatography. Enzyme Microb. Technol. 33, 445-452 https://doi.org/10.1016/S0141-0229(03)00143-1
  10. 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
  11. Kho, C.L., W.S. Tan, and K. Yusoff. 2001. Production of the nucleocapsid protein of Newcastle disease virus in Escherichia coli and its assembly into ring- and nucleocapsid-like particles. J. Microbiol. 39, 293-299
  12. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685 https://doi.org/10.1038/227680a0
  13. Lee, S.M. 1989. The primary stages of protein recovery. J. Biotechnol. 11, 103-118 https://doi.org/10.1016/0168-1656(89)90113-2
  14. Ling, T.C. 2002. Ph. D. thesis. The University of Birmingham, UK
  15. Ling, T.C., C.K. Long, W.S. Tan, B.T. Tey, A. Ariff, W.M. Wan Abdullah. 2004. Purification of filamentous bacteriophage M13 by expanded bed anion exchange chromatography. J. Micorbiol. 42, 22-232
  16. Noppe, W., I. Hanssens, and M.D. Cuyper. 1996. Simple two step procedure for the preparation of highly active pure equine milk lysozyme. J. Chromatogr. A 719, 327-331 https://doi.org/10.1016/0021-9673(95)00677-X
  17. Rabu, A., W.S. Tan, C.L. Kho, A.R. Omar, and K. Yusoff. 2002. Chimeric Newcastle disease virus nucleocapsid with parts of its viral haemagglutinin-neuraminidase and fusion proteins. Acta Virol. 46, 211-217
  18. Sumi, A., K. Okuyama, K. Kobayashi, W. Ohtani, T. Ohmura, and K. Yokoyama. 1999. Purification of recombinant human serum albumin efficient purification using STREAMLINE. Bioseparation 8, 195-200 https://doi.org/10.1023/A:1008081314112
  19. Tey, B.T., K.H.Yong, H.P. Ong, T.C. Ling, S.T. Ong, Y.P. Tan, A. Ariff, and W.S. Tan. 2004. Optimal conditions for Hepatitis B core Antigen production in shaked flask fermentation. Biotechnol Bioprocess Eng. 9, 374-378 https://doi.org/10.1007/BF02933060
  20. Thommes, J., A. Bader, M. Halfar, A. Karau, and 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
  21. Thommes, J., M. Halfar, S. Lenz, 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
  22. Ueda, E.K.M., P.W. Gout, and L. Morganti. 2003. Current and prospective applications of metal ion-protein binding. J. Chromatogr. A 988, 1-23 https://doi.org/10.1016/S0021-9673(02)02057-5
  23. Willoughby, N.A., T. Kirschner, M.P. Smith, R. Hjorth, and N.J. Titchener-Hooker. 1999. Immobilized metal affinity chromatography purification of alcohol dehydrogenase from Baker's yeast using an expanded bed adsorption system. J. Chromatogr. A 840, 195-204 https://doi.org/10.1016/S0021-9673(99)00188-0
  24. Xiang, H., R. Wynn, L-H. T. Nguyen, O.H. Ross, D.P. Ahrens, K.T. O'Neil, G.F. Hollis, and D.R. Patrick. 2002. Immobilized metal-ion affinity chromatography of recombinant Fab protein OPG C11 in the presence of EDTA-Mg (II). J. Chromatogr. A 978, 153-164 https://doi.org/10.1016/S0021-9673(02)01429-2
  25. Yusoff, K. and W.S. Tan. 2001. Newcastle disease virus: macromolecules and opportunities. Avian Pathol. 30, 439-455 https://doi.org/10.1080/03079450120078626