Production of Bovine Herpesvirus-1 Vaccine Strains in MDBK Cells Using BelloCell Bioreactor
Lekshmi S. Rajan
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Aman Kamboj
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Bikash R. Prusty
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Tapan Palai
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
L. R. Ananthakrishna
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Mohini Saini
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
Praveen K. Gupta *
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
*Author to whom correspondence should be addressed.
Abstract
Aim: To produce bovine herpesvirus-1 (BoHV-1) vaccine strains in MDBK cells using an oscillating bioreactor, BelloCell.
Place and Duration of Study: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar-243122, India, between November 2013 and June 2015.
Methodology: MDBK cells were cultured in BelloCell bioreactor seeded with MDBK cells. After 6 days incubation at 37°C under 5% CO2 tension, the MDBK cells were infected with 0.001 multiplicity of infection (MOI) of either wild-type (wt)-BoHV-1 or glycoprotein E deleted (∆gE)-BoHV-1vaccine strains and incubated further for 96 h. To analyse the vaccine potential of the harvests, inactivated wt-BoHV-1 and ∆gE-BoHV-1 were inoculated in guinea pigs. Sera from immunized guinea pigs were collected at 90 days after primary immunization and analysed for virus neutralizing (VN) antibody response specific to wt-BoHV-1.
Results: Using BelloCell bioreactor, the MDBK cell density reached to a maximum number of 1.5 x 109 to 2.0 x 109 cells after 6 days cultivation. After infection with wt-BoHV-1 and ∆gE-BoHV-1 strains, the total virus yield at 96 h post-infection was 0.5 x 1011.25 and 0.5 x 1011.83, respectively. There was no significant difference in VN antibody response in sera collected from immunized guinea pigs.
Conclusion: This study indicated that BelloCell bioreactor can be used as a simple system for high population density cell culture and large scale production of vaccines.
Keywords: Bioreactor, BelloCell, MDBK cells, bovine herpesvisus-1, immune response, guinea pig