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.)
  • Published : 1999.08.01

Abstract

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.

Keywords

References

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