PEI转染试剂

Reagent :PEIpro®. Concentration :1 mg/mL. Molecule delivered :Plasmid DNA and co-delivery of several plasmid DNA. Applications :Large-scale virus production. Virus types :Mainly adenovirus, Adeno-Associated Viruses (AAV), Lentivirus, influenza virus, retrovirus, Virus-like particles (VLP). Cell types :Mammalian producer cell lines (HEK-293, HEK-293 derivatives, BHK, VERO cell lines, virus-specific packaging cell lines. Number of transfections :100 mL of PEIpro® transfection reagent is sufficient to transfect on average 50 L of cell culture. Storage : Store PEIpro® at 5 °C ± 3°C. Expiry date is indicated in the certificate of analysis (available in "My account") and on the product. Provided with : Protocol. Certificate of Analysis. Certificate of Origin. Non-Hazardous Product Statement. Summary: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.

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 for lentivirus and AAV viral vectors(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 under the Bibliography tab.

Table 1. PEIpro^® is the reagent of choice for virus production runs in most adherent and suspension cell culture systems. Irrespective of the cell culture-based system and production scale, PEIpro^® leads to efficient lentivirus and AAV^* titer yields (*to reach highest AAV titer yields in suspension cell culture systems, we recommend to use FectoVIR^®-AAV transfection reagent).

 

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. Depending on the type of viral vector produced in suspension cell systems, the transfection reagent is critical to reach high viral vector yields. For lentivirus viral vectors produced in suspension cell sytems, high virus production yields can be reached by transfecting suspension-adapted HEK-293T cells with PEIpro^® in BalanCD^® HEK293 (Irvine Scientific) (Fig. 1).

Fig. 1: Lentivirus production in HEK-293T cells grown in suspension in BalanCD^® HEK293 (Irvine Scientific^®). HEK-293T 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^®). For transfection, a set of four plasmids was used. Lentiviral titers were measured 48 and 72 hours post-transfection (Data kindly provided by Genethon).