Particularly in the fields of iPS cells and RNAi where high efficiency gene transfer is critical, lentiviral vectors are increasingly the gene transfer system of choice for non-dividing primary and other difficult-to-transfect cells. Allele´ HiTiter™ Lentivirus systems (US patent 6,620,595) provide high titer and improved safety due to some of the most advanced and unique features:

• Further removed overlapping sequences between transfer and packaging plasmids
• Enhanced poly(A) signal to increase titer and gene expression, and prevents read-through transcription of neighboring genes
• Improved packaging of plasmids to increase packaging efficiency and to reduce the likelihood of recombination between vector components

Retroviruses are considered the easiest and fastest means to deliver genes stably to mammalian cells. Allele has upgraded retrovirus packaging from the previously offered Phoenix packaging system. The system provided either an Ecotropic packaging system (for dividing murine or rat cells) or an Amphotropic system (for dividing cells of most mammalian species, including human). We have upgraded our custom packaging method to a more efficient system that routinely result in a titer of 10⁸ or above. Allele has stopped offering the previously known Phoenix ampho and eco cells, A similar Ecotropic and Amphotopic packaging cell lines , Phoenix2, originated from different sources are currently available

• mWasabi green fluorescent protein as transfection marker
• Integrated Allele's engineered shRNA expression cassette for the highest efficiency of RNAi (US patents 7,294,504 and 7,422,896)

The baculovirus expression system (BVES) provides a high-yield and low-cost platform for expressing functional eukaryotic proteins. Most eukaryote-specific post-translational modifications, such as signal cleavage, phosphorylation, glycosylation, and disulfide bond formation, can occur in baculovirus-infected insect cells. Allele Sapphire™ Baculovirus DNA was specifically designed for efficient recombination with a polyhedrin promoter-based transfer vector by a simple co-transfection step.

• Virtually 100% recombination efficiency, which eliminates the time-consuming plaque purification step
• A protein disulfide isomerase gene was inserted at the p10 locus of the virus to ensure proper protein disulfide bond formation
• The Sapphire™ Baculovirus DNA is compatible with most of the commercial polyhedrin-based transfer vectors

Mammalian proteins may be modified differently in both insect cells and mammalian cells, for example, with different patterns of glycosylation. It has been well established that baculovirus can infect mammalian cells with broad tropism. Additionally, baculoviruses are safe for regular laboratory handling because they do not recombine with any human pathogenic viruses. The Allele Baculovirus for Mammalian Cells (BacMam) System uses the baculovirus genomic DNA with mammalian promoters and tropism enhancing factors for highly efficient protein expression of mammalian proteins.

• Enhanced tropism for broad range of mammalian cells
• Optional mWasabi green fluorescent protein and other FP markers
• A platform technology available for custom projects

Allele Biotech provides custom retrovirus or lentivirus packaging services using proprietary technologies that are unique and highly efficient, yielding 10⁸ to 10⁹ TU/ml, without any concentrating steps (US patents 6,207,455 and 6,531,123). These technologies and optimal operation procedures enable Allele Biotech to package viruses at < 1% of the market price in certain categories on per million particle basis.

• Consistently high titer without concentrating steps
• Even higher titers are provided for in vivo injection studies
• Much lower prices than other providers
• Flexible packaging sizes and multiple project discounts

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More than a decade ago, Allele Biotech staff offered the first commercial baculovirus protein expression services (under Orbigen brand) to address researchers´ unmet gene expression needs. The team now provides services at all stages of the protein expression, production, and purification processes from gene cloning to large-scale fermentation using Allele’s Sapphire BVES.

• Experience with hundreds of proteins over 10 years
• Modulated and flexible pricing scales
• Professional consultation to customers
• Timely report and advice at each stage of the project

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