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Virus production - PEIpro®

Virus production - PEIpro®

PEIpro® is a ready to use 1 mg/ml optimised linear polyethylenimine (PEI) reagent for the production of virus & recombinant proteins by Transient Gene Expression (TGE), either in adherent cells in presence of serum or in suspension HEK-293 and CHO cells grown in low serum or serum-free media.

  • Optimized for Large Scale Transient Gene Expression for virus production
  • Increased viral titers compared to other PEIs
  • Perfectly suitable for the development of bioproduction processes
  • Free of animal-origin components
  • Well characterized and lot release tested linear PEI
  • Reliable and secure supply
  • Can also be used for large scale protein production

PEIpro® undergoes full chemical testing and is compliant with the regulatory guidelines for raw materials used in bioproduction. The reagent is supplied with advanced quality controls including a specific transfection efficiency test that ensures excellent lot-to-lot consistency, and an implied license statement.

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Description

With its unique property to tightly condense several different plasmids, PEIpro® is ideal for the co-transfection of multiple plasmids that is often required for virus production.
Transfection with PEIpro® ensures high viral titers in most commonly used virus producing and packaging cell lines (HEK-293, VERO, CAP, CAP T, etc.). PEIpro® is well suited for production of all types of viruses, achieving for example viral titers > 107 IG/ml for lentiviruses and > 1011 to 1012 VG/ml for AAV.

Lentivirus and AAV production in HEK-293T and HEK-293 cells grown in suspension in BalanCD ® HEK293 (Irvine Scientific ®). HEK-293T (lentivirus) and HEK-293 (AAV) cells were thawed directly into each medium and passaged every 3 to 4 days before going into a 2L benchtop bioreactor. Cells were seeded and cultured for 3 days before being transfected by PEIpro ® (Polyplus-transfection ®). For transfection, four plasmids were used for lentivirus and three plasmids were used for AAV. Lentiviral and AAV titer were measured 48 and 72 hours post transfection (Data kindly provided by Genethon).

Compared to CaPO4, PEIpro® is more robust with transfection efficiencies ranging from 70 to 90% both in adherent and in suspension cells. It is also more powerful as it gives higher viral titers using lower DNA amount.

PEIpro ® gives higher virus titers than CaPO 4. Lentiviruses were produced in adherent HEK-293 cells grown in serum-free culture medium, using 15 µg DNA and 30 µl PEIpro ® per 75 cm 2 flask. Viral titers were determined by flow cytometry of supernatants 48 h after transfection.

With PEIpro®, transfection processes are easy to scale up. PEIpro® can be used from R&D to large scale virus or protein production. With a given 2 year shelf life and a ready to use formulation, PEIpro® is the reagent of choice for virus production runs in most cell culture media and format (Table 1) using both adherent and suspension cells.

Virus yields using PEIpro ® in different culture media for HEK-293 cells.

Examples of citations

Melidoni, A. N., Dyson, M. R., McCafferty, J. (2016). Selection of Antibodies Interfering with Cell Surface Receptor Signaling Using Embryonic Stem Cell Differentiation. Methods Mol Biol 1341, 111-32.

Stockinger, L. W., Eide, K. B., Dybvik, A. I., Sletta, H., Varum, K. M., Eijsink, V. G., Tondervik, A., Sorlie, M. (2015). The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. Biochim Biophys Acta 1854, 1494-501.

Bodnar, E., Raymond, C., Lopez, P. G., Villacres, C., Butler, M., Schoenhofen, I. C., Durocher, Y., Perreault, H. (2015). Mass spectrometric analysis of products of metabolic glycan engineering with azido-modification of sialic acids. Anal Bioanal Chem 407, 8945-58.

Li, S. K., Abbas, A. K., Solomon, L. A., Groux, G. M., DeKoter, R. P. (2015). Nfkb1 activation by the E26 transformation-specific transcription factors PU.1 and Spi-B promotes Toll-like receptor-mediated splenic B cell proliferation. Mol Cell Biol 35, 1619-32.