Technologies > VE-BEVS
Baculovirus Expression Vector System
The baculovirus expression vector system (BEVS) is a eukaryotic protein expression system that utilizes a modified insect virus for expression of recombinant proteins in insect cells. This system has several advantages over recombinant protein expression in bacterial and yeast systems.
These advantages include :
- Higher expression levels through the utilization of the polyhedrin or p10 promoter.
- Eukaryotic expression supports post-translational modifications.
- The BEVS enables simultaneous expression of multiple genes.
- Expressed proteins do not have size limitations
- The BEVS is capable of expressing cytotoxic proteins
- Safety
Vankyrin enhanced baculovirus expression vector system
ParaTechs Corp. VE-BEVS (Vankyrin enhanced baculovirus expression vector system) technology is a proprietary enhancement of the existing BEVS technology that results in a marked improvement in protein expression levels while reducing the cost of labor and materials. Cells infected with BEVS expressing a vankyrin protein from the Campoletis sonorensis ichnovirus show increased longevity and enhanced levels of recombinant protein expression that can significantly enhance heterologous protein production. Vankyrin-enhanced cell lines (VE) are genetically engineered Sf9 cells that contain a stably integrated vankyrin gene and stably express vankyrin protein. The VE-cell lines allow for enhanced protein expression from the BEVS (up to 15 fold) by providing the vankyrin protein in trans. ParaTechs currently offers three VE- cell lines for retail: 10010 (previously VE-CL-01), 10020 (previously VE-CL-02) and 10030 (previously VE-CL-03).
These cell lines show a significant enhancement in protein expression from the BEVS over regular Sf9 cell lines (Figure 1).
Figure 1. Yellow Fluorescence Protein (YFP) expression is up to 15-fold enhanced when expressed from Paratechs’ VE-BEVS. Top row shows fluorescence microscopic images of insect Sf9 cells infected with conventional BEVS expressing YFP taken 2-7d post infection (p.i.). Bottom row shows fluorescence microscopic images of insect Sf9 cells infected with Paratechs’ VE-BEVS expressing YFP taken 2-7d p.i. (20x magnification).
ParaTechs Corp. is in the final testing stage for VE-transfer vectors (VE-BEVS) that will allow for cloning and traditional expression of heterologous recombinant proteins under the polyhedrin promoter. When recombined with traditional polyhedrin locus based baculovirus systems (i.e. Baculogold, Bac-n-Blue) the vankyrin protein is expressed in cys with the recombinant protein cloned into the transfer vector. Co-expression of vankyrin protein increases recombinant protein expression up to 15-fold (Figure 2). These vectors have an expected release date of Summer 2008.
Figure 2. ParaTechs’ VE-cell lines enhance recombinant protein production up to 15-fold. Insect Sf9 cells and ParaTechs’ VE-cell lines 01, 02, and 03 were infected with baculovirus expressing a secreted eukaryotic protein (rProtein). Samples were collected after 3, 4, 5, 6 and 7 d and 2µl sample were analyzed by Western blotting. Protein production was quantified using a fluoroskan imaging system. The bar graph expresses the fold-increase in recombinant protein production of ParaTechs’ VE-cell line compared to regular Sf9 cells at the indicated time post baculovirus infection.
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