Generation of baculovirus vectors for the high-throughput production of proteins in insect cells

Robert D. Possee, Richard B. Hitchman, Kevin S. Richards, Susan G. Mann, Evangelia Siaterli, Clare P. Nixon, Helen Irving, Rene Assenberg, David Alderton, Raymond J. Owens, Linda A. King

Research output: Contribution to journalArticlepeer-review

52 Citations (Scopus)

Abstract

The baculovirus expression system is one of the most popular methods used for the production of recombinant proteins but has several complex steps which have proved inherently difficult to adapt to a multi-parallel process. We have developed a bacmid vector that does not require any form of selection pressure to separate recombinant virus from non-recombinant parental virus. The method relies on homologous recombination in insect cells between a transfer vector containing a gene to be expressed and a replication-deficient bacmid. The target gene replaces a bacterial replicon at the polyhedrin loci, simultaneously restoring a virus gene essential for replication. Therefore, only recombinant virus can replicate facilitating the rapid production of multiple recombinant viruses on automated platforms in a one-step procedure. Using this vector allowed us to automate the generation of multiple recombinant viruses with a robotic liquid handler and then rapidly screen infected insect cell supernatant for the presence of secreted proteins.

Original languageEnglish
Pages (from-to)1115-1122
Number of pages8
JournalBiotechnology and Bioengineering
Volume101
Issue number6
DOIs
Publication statusPublished - 15 Dec 2008
Externally publishedYes

Keywords

  • Automated
  • Bacmid
  • Baculovirus expression system
  • High throughput
  • Robotic liquid handler
  • Secreted protein

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