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J. Microbiol. Biotechnol. 2004; 14(1): 35-40

Published online February 28, 2004

Copyright © The Korean Society for Microbiology and Biotechnology.

Large-Scale Production of Rotavirus VLP as Vaccine Candidate Using Baculovirus Expression Vector System (BEVS)

Park Jin Yong , Hun Kim , Hi Ku Hwang 1, Hyun Sung Kim 2, Byung Ki Hur 3, Yeon Woo Ryu 3, Chang Nam An 3, Jong Soo Kim 3* and Su Jeen Lee 4*

Vaccine Research Institute GreenCross Vaccine Corp. 227-3 Gugal-Ri Yongin-City 449-903 Korea Department of Molecular Science and Technology Ajou University Suwon 442-749 Korea, 1Vaccine Research Institute GreenCross Vaccine Corp. 227-3 Gugal-Ri Yongin-City 449-903 Korea, 2Department of Biotechnology and Bioengineering Inha University Incheon 402-751 Korea, 3Department of Molecular Science and Technology Ajou University Suwon 442-749 Korea, 4Vaccine Research Institute GreenCross Vaccine Corp. 227-3 Gugal-Ri Yongin-City 449-903 Korea Department of Biotechnology and Bioengineering Inha University Incheon 402-751 Korea

Abstract

Rotavirus virus-like particle (VLP) composed of VP2, VP6, and VP7 was expressed in the Baculovirus Expression Vector System (BEVS). Sf9 cell, a host of the baculovirus, was cultured from a 0.5-1 spinner flask to the 50-1 bioreactor system. Sf9 cell was maintained at cell density between 3.0E+05 and 3.0E+06 cells/ml and grew up to 1.12E+07 cells/ml in the bioreactor. Growth kinetics was compared under different culture systems and showed similar growth kinetics with 20.1-25.2 h of doubling time. Early exponentially growing cell culture was infected with three recombinant baculoviruses expressing VP2, VP6, and VP7 protein at 1.0, 2.0, and 0.2 moi, respectively. The expression of rotavirus proteins was confirmed by Western blot analysis and its three-layered virus-like structure was observed under an electron microscope. Rotavirus VLP was semipurified and immunized in ICR mice intramuscularly. Rotavirus-specific serum antibody was detected from 2 weeks after the immunization and lasted at least 21 weeks of the post-immunization, indicating its possible use as a vaccine candidate.

Keywords: Rotavirus,virus-likeparticle,baculovirusexpressionvectorsystem,vaccine,Sf9insectcell