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flashBAC™
by Oxford Expression Technologies Ltd
Oxford Expression Technologies Ltd
• Founded in 2007 as a spin out from Oxford Brookes University
and the Natural Environment Research Council
• Technology based on insect baculovirus expression vectors
(BEVs)
• Produces high levels of recombinant proteins in insect cells
• Ideal for producing complex, highly processed proteins and
also virus-like particles (VLPs)
• Baculovirus vectors also used to transduce mammalian cells
to express genes under (eg.) CMV promoter
– The virus does not replicate in any mammalian cells
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The flashBAC™ system
(production of recombinant baculoviruses)
flashBAC™ is based on homologous recombination between a replication
deficient virus genome and a transfer vector with a foreign gene, which
also rescues virus infectivity.
• Only the recombinant virus with a restored ORF 1629 can replicate
• No need to plaque purify virus
• Represents a ONE-STEP process that can be used in high or low throughput
applications
flashBAC™ saves you time, simplifies the expression process and increases
protein yield compared to other systems, as shown in the following slides
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flashBAC™ time line comparison with
other baculovirus systems
One step
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Comparison of kinase production levels in
flashBAC GOLD™ and Bac-to-Bac®
expression systems
®
The Western blots show a significant increase in kinase levels produced in the flashBAC GOLD™ system
compared to Bac-to-Bac ®, at each time point recorded. This demonstrates that flashBAC GOLD™ is a
favourable expression system when expressing certain proteins. (Hitchman RB el al. 2010 Improved
expression of secreted and membrane-targeted proteins in insect cells doi: 10.1042/BA20090130.)
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flashBAC™ variants
(Available in kits for own use)
• flashBAC™
– Original, good for general purpose expression and BacMAM use
• flashBAC GOLD™
– Chitinase, cathepsin genes deleted, ideal for protein secretion
• flashBAC ULTRA™
– Also lacks p10 gene and other non-essential genes to enhance cell
stability late in infection
• flashBAC PRIME™
– Essentially wild type genome, but superior for producing some VLP’s
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flashBAC ULTRA™
EGFP
Comparison of EGFP expression between the
different flashBAC™ platforms. flashBAC ULTRA™
produces significantly higher yields of EGFP.
(Hitchman et al. 2010, Genetic modification of a
baculovirus vector for increased
expression in insect cells, DOI 10.1007/s10565009-9133-y)
flashBAC ULTRA™ has additional
deletions to the non-essential genes
p26, p10 and p74. These deletions
increase cell stability and lower the
burden on virus-infected cells,
allowing more target protein to be
produced.
This combination makes flashBAC
ULTRA™ the best platform for
expression of membrane and
cytoplasm targeted proteins.
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Advantages in flashBAC™ in human
therapeutics and vaccines
• Therapueutics and flashBAC™ - Use of BEVs in
human therapeutics and vaccine work is set to
expand greatly after the approval of baculovirusderived Cervarix and Flublok, HPV and influenza
vaccines (respectively).
• flashBAC™ uses homologous recombination unlike
E.coli based systems (Bac-to-Bac®), which can retain
residual antibiotic resistance markers and bacterial
sequences, flashBAC™ would therefore be first
choice for any therapeutic based protein production.
(Kwang et al. 2016 Manufacturing of AcMNPV baculovirus vectors to enable gene
therapy trials, Nature. doi:10.1038/mtm.2015.50)
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Summary
• flashBAC™ is a one-step process that:
-
Saves you time
Saves equipment
Simplifies protein expression
Decreases the chance of handling errors
• flashBAC™ has 4 variant platforms to suit your needs and
optimise the expression levels of your protein
• flashBAC™ can be used in mammalian cell lines using
BacMAM technology, ideal for producing virus-like
particles (VLPs)
• Take advantage of the Gateway™ technology with our
pOET Gateway™ transfer vectors
• OET is a name that you can trust for all things baculovirus
Apply now for a free transfer vector
Once you start using our flashBAC ULTRA™
system, we know you’ll keep using it.
Contact OET for a free 3 reaction flashBAC
ULTRA™ kit when you purchase any pOET
transfer vectors!
Contact us on (00 44) 0 1865 483236 or at [email protected]
Checkout our website www.oetltd.com