Chapter 8
Bioreactor-Based Antigen Production Process Using
the Baculovirus Expression Vector System
Julie Harnischfeger, Lukas K € aßer, Jan Zitzmann, Denise Salzig,
and Peter Czermak
Abstract
Several vaccines are already produced using the baculovirus expression vector system (BEVS). This chapter
describes methods for generating recombinant baculoviral DNA (also called bacmid) for cultivating
Spodoptera frugiperda Sf-9 cells and producing a baculovirus stock from the recombinant bacmid and for
producing a protein-based vaccine with the BEVS in a stirred tank reactor.
Key words Process analytical technology, Online process monitoring, Dielectric spectroscopy, Spodoptera frugiperda, Sf-9 cells, Antigen production, Cell cultivation
1 Introduction
Various competing production platforms, based on different host
organisms, are involved in the production of vaccines. Within this
field, the baculovirus expression vector system (BEVS) is, although
established more than 30 years ago, just becoming a state-of-theart technology. Several BEVS-based vaccines for veterinary and
human use, which are based on protein subunits (e.g., Flublock
® ,
Protein Sciences Corporation, human influenza vaccine) or virus
like particles (e.g., Cervarix
® , GSK, human papillomavirus vaccine),
have been approved and marketed in the past decades [1]. Consequently, BEVS can be considered to be an established tool, not only
for research purposes but also for industrial production. The BEVS
technology itself is based on insect cell-lines as such as Sf-9 or Sf-21
(Spodoptera frugiperda) and a corresponding recombinant Autographa californica nuclear polyhedrosis virus (AcNPV), carrying
the genetic information for the target vaccine to be expressed.
Because the production is based on virus infection, rather than on
a stable chromosomal integration of the foreign genetic information, BEVS offers manufacturing speed, high flexibility, and yields
Blaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_8, © Springer Science+Business Media, LLC, part of Springer Nature 2021
95
Bioreactor-Based Antigen Production Process Using
the Baculovirus Expression Vector System
Julie Harnischfeger, Lukas K € aßer, Jan Zitzmann, Denise Salzig,
and Peter Czermak
Abstract
Several vaccines are already produced using the baculovirus expression vector system (BEVS). This chapter
describes methods for generating recombinant baculoviral DNA (also called bacmid) for cultivating
Spodoptera frugiperda Sf-9 cells and producing a baculovirus stock from the recombinant bacmid and for
producing a protein-based vaccine with the BEVS in a stirred tank reactor.
Key words Process analytical technology, Online process monitoring, Dielectric spectroscopy, Spodoptera frugiperda, Sf-9 cells, Antigen production, Cell cultivation
1 Introduction
Various competing production platforms, based on different host
organisms, are involved in the production of vaccines. Within this
field, the baculovirus expression vector system (BEVS) is, although
established more than 30 years ago, just becoming a state-of-theart technology. Several BEVS-based vaccines for veterinary and
human use, which are based on protein subunits (e.g., Flublock
® ,
Protein Sciences Corporation, human influenza vaccine) or virus
like particles (e.g., Cervarix
® , GSK, human papillomavirus vaccine),
have been approved and marketed in the past decades [1]. Consequently, BEVS can be considered to be an established tool, not only
for research purposes but also for industrial production. The BEVS
technology itself is based on insect cell-lines as such as Sf-9 or Sf-21
(Spodoptera frugiperda) and a corresponding recombinant Autographa californica nuclear polyhedrosis virus (AcNPV), carrying
the genetic information for the target vaccine to be expressed.
Because the production is based on virus infection, rather than on
a stable chromosomal integration of the foreign genetic information, BEVS offers manufacturing speed, high flexibility, and yields
Blaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_8, © Springer Science+Business Media, LLC, part of Springer Nature 2021
95