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

Park, Jin-Yong (Vaccine Research Institute, GreenCross Vaccine Corp., Department of Molecular Science and Technology, Ajou University)
Kim, Hun (Vaccine Research Institute, GreenCross Vaccine Corp., Department of Molecular Science and Technology, Ajou University)
Hwang, Hi-Ku (Vaccine Research Institute, GreenCross Vaccine Corp.)
Lee, Su-Jeen (Vaccine Research Institute, GreenCross Vaccine Corp.)
Kim, Hyun-Sung (Vaccine Research Institute, GreenCross Vaccine Corp., Department of Biotechnology and Bioengineering, Inha University)
Hur, Byung-Ki (Department of Biotechnology and Bioengineering, Inha University)
Ryu, Yeon-Woo (Department of Molecular Science and Technology, Ajou University)
An, Chang-Nam (Vaccine Research Institute, GreenCross Vaccine Corp.)
Kim, Jong-Soo (Vaccine Research Institute, GreenCross Vaccine Corp.)
Publication Information
Journal of Microbiology and Biotechnology / v.14, no.1, 2004 , pp. 35-40 More about this Journal
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-like particle; baculovirus expression vector system; vaccine; Sf9 insect cell;
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