PEIpro®

Large-scale Virus Production

  • 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

Specifications

Reagent

PEIpro®

Molecule delivered

DNA

Applications

Virus production
Protein production

Cell types

Adherent HEK-293 and derivatives grown in serum containing medium
Suspension HEK-293 and derivatives grown in synthetic serum-free medium
Other cell lines used for virus production such as VERO, CHO and BHK cells

Number of transfections

100 mL of PEIpro® transfection reagent is sufficient to transfect on average 50 L of cell culture

Storage

4 °C for up to 2 years

Summary

Focus on your production yield, use a PEI you can rely on!

PEIpro® is a DNA transfection reagent for production of viruses, recombinant proteins and antibodies either in adherent cells grown in presence of serum or in suspension HEK-293 and CHO cells grown in low serum or serum-free media.

PEIpro® benefits from product and process improvements in terms of manufacturing process, product characterization and proven transfection efficiency for large-scale virus and protein production. Combined with appropriate and advanced quality controls as well as lot release testing, PEIpro® enables biopharmaceutical companies to develop consistent, reliable and secure processes for reproducible production of viruses, recombinant proteins and antibodies of the highest quality. For production of clinical batches of virus, Polyplus-transfection® offers a highly qualified grade of PEIpro®, PEIpro®-HQ.

File icone Read the article published in GEN: Simplifying the Efficient Clinical-Grade Production of Viruses

Ordering information

Catalog NumberAmount of reagent
115-01010 ml
115-100100 ml
115-4004 x 100 ml

Large sizes are available upon request.

Request a quote

Optimized for transfection compared to other PEIs

The linear form of PEIpro® and the manufacturing process developed by Polyplus-transfection® ensure a high, stable and reproducible amount of protonable amines available for transfection while providing a truly deacylated molecule and an extremely low polymer chain length variation. As a result, PEIpro® is more powerful as it requires less DNA and reagent than any other linear PEI (Fig. 1) for increased virus and protein production yield in HEK-293 and CHO cells grown in serum free media.

PEIpro - Comparison PEIs
Fig. 1: PEIpro® requires less reagent and similar to lower DNA amount compared to other PEIs. Suspension HEK-293 cells were seeded at 1×106 cells/ml in serum free medium and transfected with PEIpro®, PEI “Max” and L-PEI 25 kDa (Polysciences, Warrington, PA) resuspended at 1 mg/ml. Luciferase expression was assayed 48 h after transfection using a conventional luciferase assay.

PEIpro® has been formulated at 1 mg/ml to fit current standards for transfection with PEI. This concentration simplifies the transfection protocol optimization. Please Contact the Technical Support for any questions about the protocol.

Well characterized with lot release testing

PEIpro® is fully synthetic and is free of components of animal-origin.

The quality of PEIpro® is continuously assessed during the manufacturing process with appropriate control testing. Systematic lot management and release testing is performed for each lot produced. A standardized transfection assay is performed with suspension HEK-293 cell line in serum-free media to ensure the reliability and reproducibility of the transfection efficiency and protein yields under conditions that are suitable for Biomanufacturing.

PEIpro® undergoes stringent quality controls to ensure complete sterility, absence of mycoplasma and very low level of bacterial endotoxins following pharmacopoeia standards and is supplied with adequate documentation. A full list of QCs is available upon request (Contact the Technical Support). For production of clinical batches of virus and protein, Polyplus-transfection® offers a highly qualified grade of PEIpro®, PEIpro®-HQ.

Great lot to lot consistency

PEIpro® is manufactured and formulated using a highly controlled and reproducible production process. By assuring reproducibility from one run to the next, irrespective of the operator, PEIpro® greatly facilitates process standardization; a key advantage for large-scale virus and protein production. Moreover, PEIpro® is released using advanced quality controls including a specification for transfection efficiency that guarantees excellent lot-to-lot consistency (Fig. 2).

PEIpro - reproductibility IgG3
Fig. 2: Excellent lot-to-lot protein yield reproducibility using PEIpro®. Suspension HEK-293 cells were seeded at 1×106 cells/ml in serum-free medium and transfected with PEIpro® following the standard protocol. IgG3-Fc production was assayed 48 h after transfection using protein G affinity quantification (HPLC).

Versatile and easy to scale up

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.

PEI - Virus yields compared with different culture media
Table 1: Virus yields using PEIpro® in different culture media for HEK-293 cells.

Virus production

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 (Fig. 1).

PEIpro - Lentivirus and AAV production
Fig. 1: 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 (Fig. 2.).

PEIpro - Virus titers
Fig. 2: PEIpro® gives higher virus titers than CaPO4. Lentiviruses were produced in adherent HEK-293 cells grown in serum-free culture medium, using 15 µg DNA and 30 µl PEIpro® per 75 cm2 flask. Viral titers were determined by flow cytometry of supernatants 48 h after transfection.

PEIpro® is particularly suited for virus production using FreeStyle™ F17 medium (Life Technologies™).

Read more…

Protein production

PEIpro® is perfectly suited for large scale protein production in both HEK-293 and CHO cells. Indeed, this reagent is compatible with various media formulations for both CHO and HEK-293 cells.

Read more…

Here is a selection of relevant references using PEIpro®, more are available in our Polyplus-transfection Database.

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.

Fuller, J. P., Stavenhagen, J. B., Christensen, S., Kartberg, F., Glennie, M. J., Teeling, J. L. (2015). Comparing the efficacy and neuroinflammatory potential of three anti-abeta antibodies. Acta Neuropathol 130, 699-711.

 

Genethon proudly uses PEIpro for production of pre-clinical batches of Lentivirus & AAV.

Genethon Logo

PEIpro® is fully synthetic and free of any animal-origin component. The quality of PEIpro® is continuously assessed and documented during manufacturing with the appropriate control testings. Further systematic lot management and release testing is performed for each lot produced. A qualified transfection assay ensures reliability and reproducibility of the transfection efficiency under conditions suitable for biomanufacturing. PEIpro® undergoes sterility, mycoplasma and bacterial endotoxin testing according to pharmacopoeia standards. A full list of QCs is available upon request (Contact the Technical Support).

In addition, Polyplus-transfection® is ISO 9001 Quality Management System accredited since 2002; this level of certification assures global customers that the supplier has established reliable and effective processes for product development, manufacturing, sales and customer support.