FlashBacGold is based on the OET's patented FlashBac system which uses a single-step method for the rapid and automated generation of baculovirus expression vectors
Oxford Expression Techologies (OET) chose PepTalk 2008 in San Diego, California to launch FlashBacGold, a baculovirus expression vector designed specifically to reduce proteolysis, maximise protein secretion and improve membrane protein targeting.
Plaque purification is not required.
FlashBacGold is back-compatible with all existing baculovirus transfer vectors based on homologous recombination in insect cells at the polyhedrin locus.
The main advantages of FlashBacGold is that it is rapid and simple.
The one-step procedure saves time and the method is suitable for the production of single or multiple viruses.
FlashBacGold is also very flexible and is suitable for use with manual and automated methodologies.
To improve the yield of difficult to express proteins, FlashBacGold uses a modified baculovirus genome that has had two non-essential genes deleted.
Deletion of genes encoding chitinase (chiA), an enzyme with exo- and endochitinase activity, and v-cath, a cathepsin-like cysteine protease, improves the efficacy of the secretory pathway and results in a greatly enhanced yield of recombinant proteins that are secreted or membrane targeted (in comparison to recombinant viruses that synthesise chiA and v-cath).
Results also show a significant reduction in degradation of protease-sensitive targets and increased production and stability of some intracellular proteins.
FlashBac uses a genetically engineered form of baculovirus, a virus that replicates only in the cells of butterflies, moths and caterpillars.
When moth cells are infected with this virus in culture, they produce large amounts of a given protein.
The resultant proteins are antigenically, immunogenically and functionally similar to the native protein.
The FlashCac system enables fast and simultaneous production of multiple recombinant viruses and is ideal for high-throughput and automated systems.
It allows scientists to produce proteins faster, more easily and cost-effectively.
The FlashBacGold kit contains Flashbac DNA which lacks part of an essential gene and contains a bacterial artificial chromosome (BAC).
The essential gene deletion prevents virus replication within insect cells but the BAC allows the viral DNA to be maintained and propagated.
Homologous recombination between Flashbac DNA and a transfer vector containing a gene of choice restores the function of the essential gene and simultaneously removes the BAC sequence.
The recombinant virus subsequently replicates to produce a genetically homogenous recombinant virus population without the need to perform a plaque assay.
After five days, virus can be added directly to insect cells to amplify a high titre working virus stock.
This one-step procedure for making recombinant baculoviruses greatly facilitates the process of high throughput production of baculovirus expression vectors via automated systems.
Commenting on the introduction of Flashbacgold, James Bernard, co-investor and acting CEO said, "The new system has been designed to meet the needs of academic and industrial communities who are looking for even faster, generic high-throughput protein production.
"Flashbacgold is rapid, simple and flexible and provides higher yields of secreted and membrane-targeted proteins with the minimum of effort.
"We are delighted with the positive response that we received at PepTalk 2008".
