- InVitria & Pall Biotech Posters
We are proud to feature two new posters co-authored with Pall Biotech. Each poster can be accessed by clicking on the images to the right or the hyperlinked poster titles below:
Application of Aber’s FUTURA Biomass Probes to Inform Transfection and Cell Lysis in iCELLis® Bioreactor-Based AAV Manufacturing
Utilization of classical adherent formats in large scale viral vector manufacturing can have significant setbacks due to the lack of scalability of production vessels typically used at small scale. The iCELLis fixed bed bioreactor has emerged as an enabling technology to efficiently scale adherent-based processes in a controlled and highly integrated environment. This technology has been developed for the clinical manufacture of lentiviral and adeno-associated vectors with commercially viable yields. Aber’s FUTURA Biomass probes, which induce polarization of cells and measures the resulting capacitance of the medium in pF/cm, can be employed with the iCELLis bioreactor to provide direct online information on cell biomass during a viral vector production run. Routine utilization of these probes can provide invaluable online information regarding cell growth and health, control of feed/perfusion rate, and the identification of optimal time for transfection or harvest. Here, we utilized Aber’s FUTURA Biomass probes in iCELLis bioreactor runs to produce an AAV-2 GFP vector in OptiPEAK HEK293t chemically defined media, using different feeding strategies and harvest protocols of the bioreactor. Data obtained from the biomass probes are correlated with overall functional vector titer to identify ideal capacitance trends that are predictive of bioreactor productivity. Taken together, these data suggest the Aber’s FUTURA Biomass probes can be used to identify ideal capacitance ranges to commence major unit operations of the manufacturing process to maximize vector yields.
Efficient Lentiviral Vector Production in a Chemically Defined, Blood-Free and Serum-Free Medium, Scalable to the iCELLis® Technology
Retroviral vectors are a promising candidate for the treatment of rare, monogenic diseases. Lentivirus — a type of retrovirus based on HIV — is currently being clinically evaluated in stage 3 trials for the treatment of rare blood disorders in addition to the genetic modification of human T cells in oncology applications. While the efficacy looks promising in the clinic, numerous questions surrounding the feasibility of large-scale manufacturing of lentivirus remain. Traditionally, production of these retroviral vectors has been performed using adherent platforms that rely on the use of fetal bovine serum for the adherence and growth of HEK cells used to produce lentivirus vectors. At scale, fetal bovine serum presents numerous problems including but not limited to lot-to-lot variation, constraints on the global supply chain, and increasing cost due to global demand. To overcome these limitations, we have developed OptiPEAK® HEK serum-free, chemically defined cell culture medium that is free from any blood-derived proteins and supports adherent HEK cells in 2D and 3D formats. With OptiPEAK HEK cell culture medium, we are able to achieve equivalent growth kinetics and viral titer compared to medium supplemented with serum. We also demonstrate that OptiPEAK HEK medium can be readily scaled up to an iCELLis Nano bioreactor, achieving high viral titers without the addition of serum. The presented data here demonstrate that high titer, retroviral vectors can be manufactured without the constrictions brought on by the inclusion of serum in cell culture medium.