Journal of Microbiology and Biotechnology

  • 제9권4호
  • /
  • Pages.386-392
  • /
  • 1999
  • /
  • 1017-7825(pISSN)
  • /
  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Large-Scale Culture of Hepatitis A Virus in Human Diploid MRC-5 Cells and Partial Purification of the Viral Antigen for Use as a Vaccine

  • Kim, Hyun-Seok (R&D Center of Pharmaceuticals, Institute of Science and Technology, Cheiljedang Inc.) ;
  • Chung, Yong-Ju (R&D Center of Pharmaceuticals, Institute of Science and Technology, Cheiljedang Inc.) ;
  • Jeon, Yeong-Joong (R&D Center of Pharmaceuticals, Institute of Science and Technology, Cheiljedang Inc.) ;
  • Lee, Sung-Hee (R&D Center of Pharmaceuticals, Institute of Science and Technology, Cheiljedang Inc.)
  • 발행 : 1999.08.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95％ of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40％ of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

키워드

참고문헌

  1. JAMA v.264 Risk factors for acute non-A, non-B hepatitis in the United States and association with hepatitis C Alter, M. J.;S. C. Hadler;F. N. Judson
  2. J. Med. Virol. v.22 Cytopathology, plaque assay, and heat inactivation of hepatitis A virus strain HM175 Anderson, D. A.
  3. Prog. Med. Virol. Basel v.37 Inactivated candidate vaccines for hepatitis A Andre, F. E.;A. Hepburn;E. D. Hondi
  4. Immunochemistry v.8 Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration Avrameas, S.;T. Ternynck
  5. J. Clin. Microbiol. v.20 Primary isolation and serial passage of Hepatitis A virus strains in primate cell cultures Binn, L. N.;S. M. Lemon;R. H. Marchwicki;R. R. Redfield;N. L. Gates;W. H. Bancroft
  6. Viral Hepatitis and Liver Diseases Inactivated hepatitis A vaccine produced in human diploid MRC-5 cells Binn, L. N.;W. H. Bancroft;K. H. Eckels;R. H. Marchwicki;D. R. Dubois;L. V. S. Burke;A. J. Zuckerman(ed.)
  7. J. Infect. Dis. v.153 Preparation of a prototype inactivated hepatitis A virus vaccine from infected cell culture Binn, L. N.;W. H. Bancroft;S. M. Lemon;R. H. Marchwicki;J. W. Leduc;C. J. Trahan;E. C. Staely;C. M. Keenan
  8. J. Virol. Methods v.47 Rapid and efficient purification of hepatitis A virus from cell culture Bishop, N. E.;D. L. Hugo;S. V. Borovec;D. A. Anderson
  9. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford, M.
  10. Intervirology v.18 Biophsical and biochemical characterization of hepatitis A virus Coulepis, A. G.;S. A. Locarnini;E. G. Westaway;G. A. Tannock;I. D. Gust
  11. J. Biol. Methods v.18 Continuous production of hepatitis A virus PLC/PRF/5 cell cultures: Use of antigen for serology Crance, J. M.;J. Passagot;E. Biziagose;R. Deloince
  12. J. Med. Virol. v.22 Development of a plaque assay for a cytopathic, rapidly replicating isolate of hepatitis A virus Cromeans, T.;M. D. Sobsey;H. A. Fields
  13. Infect. Immun. v.32 Propagation of human hepatitis A virus in African green monkey kidney cell culture: Primary isolation and scrial passage Daemer R. J.;S. M. Feinstone;I. D. Gust;R. H. Purcell
  14. Virology v.157 Replicative events in hepatitis A virus-infected cells de Chastonay, J.;G. Siegel
  15. J. Virol. Methods v.32 Large-scale purification of inactivated hepatitis A virus by centrifugation in non-ionic gradients Dubois, D. R.;K. H. Eckels;J. Ticehurst;L. N. Binn;R. L. Timchark;D. A. Barvir;C. T. Rankin;S. P. O'Neill
  16. Science v.182 Hepatitis A: Detection by immune electron microscopy of a virus-like antigen associated with acute illness Feinstone, S. M.;A. Z. Kapikian;R. H. Purcell
  17. Intervirology v.20 Taxonomic classification of hepatitis A virus Gust, I. D.;A. G. Coulpis;S. M. Feinstone;S. A. Locarnini;Y. Moritsugu;R. Najera;G. Siegel
  18. J. Infect. Dis. v.151 The origin of the HM175 strain of Hepatitis A virus Gust, I. D.;N. I. Lehmann;S. Crowe;M. McCrorie;S. A. Locarnini;C. R. Lucas
  19. Virus Res. v.12 Detection of hepatitis A virus RNA and capsid antigen in individual cells Harmon, S. A.;D. F. Summers;E. Ehrenfeld
  20. Infect. Immun. v.42 Antigenic relatedness of two strains of hepatitis A virus determined by crossneutralization Lemon, S. M.;L. N. Binn
  21. J. Virol. v.54 Infectious hepatitis A virus particles produced in cell culture consist of three distinct types with different buoyant densities in CsCl Lemon, S. M.;R. W. Jansen;J. E. Newbold
  22. Intervirology v.18 Classification of hepatitis A virus as enterovirus type 72 and hepatitis B virus as hepadnavirus typeⅠ Melnick, J. L.
  23. Archives of Virology Suppl v.2 Classification and Nomenclature of Viruses: The Fifth Reprot of the International Committee on Taxonomy of Viruses. Wien Minor, P. D.;R. I. B. Franki(ed.);C. M. Fauquet(ed.);D. L. Knudson(ed.);F. Brown(ed.)
  24. Appl. Environ. Microbiol. v.53 Production of cytopathology in FRhK-4 cells by BS-C-1 passaged hepatitis A virus Nasser, A. M.;T. G. Metcalf
  25. Vaccine v.11 Preparation and immunogenicity of an inactivated hepatitis A vaccine Pellegrini, V.;N. Fineschi;G. Matteucci;I. Marsili;L. Nencioni;M. Puddu;H. Garelick;A. J. Zuckeman
  26. Proc. Soc. Exp. Bio. Med. v.160 Propagation of human hepatitis A virus in cell culture in vitro Provost, P. J.;M. R. Hilleman
  27. Am. J. Hyg. v.27 A simple method of estimating fifty percent endpoints Reed, L. J.;H. A. Muench
  28. J. Virol. Meth. v.72 Identification and characterization of incomplete hepatitis A virus particles Ruchti, F.;G. Sicgel;M. Weitz
  29. J. Virol. v.56 Isolation and molecular cloning of a fast-growing strain of a human hepatitis A virus from its double-stranded replicative form Venuti, A.;C. D. Russo;N. Del Grosso;A. M. Patti;F. Ruggeri;P.R. De Stasio;M .G. Martinello;P. Pagnotti;A. M. Degener;M. Midulla;A. Pana;R. Perez-Bercoff