INNOPHEX On-Demand Sessions

Pall evaluated the Allegro™ STR bioreactor family for its performance as a scalable vector production platform for an investigational viral vector-based gene replacement therapy to treat Sanfilippo syndrome type A (MPS IIIA). The scalability of a PEI mediated transfection process was evaluated between the 50 L and 500 L scale for production of this AAV vector. Performance of these bioreactors were evaluated based on cell growth, metabolic profile, and vector production.

The Pall Accelerator Process Development services aim to co-develop an optimal process platform solution with customers. We leverage our industry experts, proven methods, and integrated bioprocessing equipment to reduce time to market, lower costs, and minimize risk.

Gene therapy is a rapidly expanding field with many unique challenges, particularly those associated with downstream purification. Adeno-Associated Virus (AAV) and Lentivirus are two prominent viral vectors with significant traction in the gene therapy field. Here we asses clarification, tangential flow filtration, and final sterile filtration techniques for both Suspension and Adherent cultured AAV and Lentivirus.

Significant challenges persist in Adeno-Associated Virus (AAV) purification as AAV harvests typically contain a majority population of empty capsids that generate an immune response without delivering the therapeutic payload.

Here we assess the performance of anion exchange (AEX) membrane chromatography as the polishing stage of an AAV platform proces. Utilizing a novel 1 mS/cm step gradient approach, we are able to demonstrate separation of empty and full AAV capsids of different AAV serotypes. This technique is scalable between the 0.86 mL Mustang Q XT Acrodisc® unit and 5 mL Mustang Q XT capsule, and effectively clears residual host cell protein and DNA contaminants.

A key aspect of Chemistry Manufacturing and Controls (CMC) documentation for complex biological products is the application of Quality by Design (QbD) principles: A rationale of quality being achieved by process design rather than relying on final quality testing alone. The work presented here provides a framework to illustrate the concept and initial thoughts on using QbD concepts for gene therapy, specifically Adeno-Associated Virus (AAV) production.

The number and variety of cell-therapy products is increasing, there is a need for a flexible manufacturing platform which can be implemented in research labs and easily scaled up to expedite process development, pre-clinical testing, and large-scale expansion of adherent stem and other primary cells. Pall’s Xpansion single-use bioreactor offers a tightly-controlled, scalable manufacturing platform for cell therapy applications, allowing high-density expansion of adherent cells while maintaining their functionality. This presentation will highlight a variety of interesting and new proof of concept, scale-up, and manufacturing stories using Xpansion bioreactors.

As monoclonal antibody production has become routine for drug suppliers, the desire to intensify, improve, and reduce the cost of processes has become omnipresent. With these goals in mind there appears to be a growing need for an end-to-end automated process with a robust control strategy. Here a fully automated approach for the control of a continuous downstream process has been evaluated for its ability to coordinate a process with minimal intervention from operators. Enacting this control strategy facilitated an end-to-end process for 24 hours at two scales as well as a small scale 4-day process. These processes were executed with minimal operator intervention where product quality was maintained for all processes. Some particular challenges such as volumetric throughput limitations, scale-up, equipment sizing, etc. were explored in a lab scale setting up to a 20x scale up for the Protein A capture step. By operating our downstream process, a deeper fundamental understanding of the purification platform can be developed. This operation seconds as a proving ground for the control strategy the team develops for a particular process. By operating the process, deficiencies in the control can be identified and optimized to improve the efficiency, efficacy, and product quality. Through these means further understanding of critical process parameters and how they affect the critical quality attributes at the whole process level, not just at the individual unit operation level, has been achieved. Using the knowledge gained from these studies we can identify which parameters have the largest impact on the process and which should be controlled within the tightest engineering limits from a process perspective, and further develop and implement automation improvements to eliminate control deficiencies.

Increased demand for irradiated health-care goods coupled with supply chain complexities related to gamma irradiation, have driven an industry-wide need to understand, risk assess and qualify X-ray sterilization as an equivalent alterative to supplement gamma irradiation capacity.  In addition to highlighting implementation timelines and strategies, examples of qualification data will be shown as well as a risk based approach to assessing the impact to your bioprocess.