Large-scale virus Production
- Optimized PEI-based transfection reagent for scalable virus production
- Superior viral titers compared to other commercial PEIs and Calcium Phosphate
- Suitable for Process Development of large-scale virus production
- Reliable and secure supply
Plasmid DNA and co-delivery of several plasmid DNA
Large-scale virus production
Mainly adenovirus, Adeno-Associated Viruses (AAV), Lentivirus, influenza virus, retrovirus, Virus-like particles (VLP)
Mammalian producer cell lines (HEK-293, HEK-293 derivatives, BHK, VERO cell lines, virus-specific packaging cell lines) grown in adherent or suspension cultures.
|Number of transfections|
100 mL of PEIpro® transfection reagent is sufficient to transfect on average 50 L of cell culture
5 ± 3 °C, 3 years from manufacturing date
PEIpro® transfection reagent is the leading chemical-based DNA transfection reagent that offers flexibility and scalability to develop a robust and sustainable Process Development for viral vector production. PEIpro® benefits from extensive research development in PEI polymer chemistry and formulation to achieve highest transfection efficiency in both adherent and suspension cell culture systems. Protocols established with PEIpro® during Process Development are directly implementable for large-scale manufacturing of preclinical and clinical batches of viral vectors with the higher quality grades of PEIpro®: PEIpro®-HQ and PEIpro®-GMP.
|Reference Number||Amount of reagent|
|115-040||4 x 10 ml|
|115-400||4 x 100 ml|
Optimized PEI-based transfection reagent for scalable virus production
With its unique property to efficiently condense several plasmids of different sizes, PEIpro® transfection reagent is ideal for optimal co-transfection of several plasmids containing the gene of interest and necessary viral components to produce full recombinant virions. PEIpro® has benefited from extensive developments that made it the unique PEI-based transfection reagent that can offer the flexibility and robustness that is needed during Process Development and Scale-up; it is compatible with commercially available adherent and suspension virus production systems, particularly for AAVs and lentiviruses (Table 1). Published work on AAV and lentivirus production with PEIpro®, and on other types of viruses (adenovirus, retrovirus virus-like particle (VLP)) production, are available here : PEIpro literature.
Suspension cell culture-based systems can offer considerable advantages in simplifying the scale-up process, in terms of cell culture conditions and harvesting. For this, several animal-free synthetic media are commercially available to maintain high productivity in cells lines adapted for growth in suspension. Typically, high virus production yields can be reached for both AAV and lentivirus by transfecting suspension-adapted HEK-293T cells with PEIpro® (Fig. 1).
Fixed-bed bioreactors are also an attractive scale-up model as they allow for increased volumetric productivity with low shear stress cultivation of adherent cells. Development of scalable viral vector production has been successfully implemented in the fixed-bed reactor iCELLis® (Pall) in combination with the use of PEIpro®. For example, during the scale-up production of a recombinant AAV-8 (rAAV-8) from 0.8 m2 to 4 m2 iCELLis® Nano bioreactor, the established PEIpro® transfection protocol could be readily transferred without further optimization. The virus yields obtained in the 4 m2 iCELLis® Nano bioreactor demonstrated the process scalability and robustness. For more details, see here.
Optimized for transfection compared to other commercial PEIs and Calcium Phosphate
PEIpro® vs. commercial PEIs
The extensive research development invested in PEIpro® and the manufacturing process developed by Polyplus-transfection® has made it the unique PEI-based transfection reagent that benefits from extensive research development in polymer chemistry and formulation, in terms of polymer structure, deacetylation degree, polydispersity index and molecular weight. As a result, PEIpro® is the gold reference among commercially available PEIs and requires lowest DNA amount and volume of reagent to ensure reproducibly high production yields (Fig. 2).
PEIpro® vs. Calcium Phosphate
Calcium phosphate (CaPO4) is known to be inefficient for transfection of cells grown in suspension, costly in the amount of plasmid DNA required, and inconsistent due to several fluctuating parameters during complexation (pH, calcium and phosphate concentrations). Compared to CaPO4, PEIpro® is efficient with high transfection efficiencies ranging from 70 to 90% both in adherent and in suspension cells, which correspondingly leads to higher virus titer yields (Fig. 3).
Great lot-to-lot consistency
PEIpro® is manufactured and formulated using a highly controlled and strict manufacturing process. By ensuring reproducibility from one run to the next, irrespective of the operator, PEIpro® greatly facilitates process standardization; a key advantage for large-scale production. Moreover, PEIpro® is released using advanced quality controls including a specification for transfection efficiency that guarantees excellent lot-to-lot consistency (Fig. 4).
Here is a selection of relevant references, patents and companies using PEIpro®, more are available in our Polyplus-transfection Database.
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.