Resubmission - Development and scale-up of Continuous Countercurrent Tangential Chromatography (CCTC) for next generation AAV vector manufacturing (Direct to Phase II)

PROJECT SUMMARY The overall goal of the proposed Direct to Phase II SBIR project is to commercialize Continuous Countercurrent Tangential Chromatography (CCTC) for recombinant AAV purification. This novel platform will enable column-free, continuous, and single-use purification of rAAVs. Successful commercialization of the CCTC system in this field will lead to at least a 50% reduction

in downstream production costs, increased recoveries (by 40%) and improved product quality for these life saving therapies that provide effective treatments and cures for such conditions as cancer, hemophilia and many others. Continuous and flexible single-use processing for biologics has been established as a significant

unmet need. Dr. Janet Woodcock, Director of the FDA Center for Drug Evaluation and Research, stated in a 2011 AAPS meeting: “It is predicted that manufacturing will change in the next 25-years as current manufacturing practices are abandoned in favor of cleaner, flexible, more efficient continuous manufacturing.” Significant bottlenecks in downstream processing as well

as the explosive growth in the number of gene therapies and specifically rAAVs in development are the primary drivers of this unmet need. Our aim is to commercialize CCTC for the purification of rAAV to significantly improve recovery, product quality, and process productivity when compared with status-quo column

chromatography. CCTC provides a steady state purification platform with >10x reduction in cycle time and greater productivity compared to current column-based purification. Preliminary data obtained with a commercial rAAV product show at least a 5-15x increase in productivity and 15-20% improvement in recovery for a capture operation. In addition, because the

CCTC platform runs at low pressure, small particle size resins may be used to significantly increase binding capacity and resolution. This feature could enable finely tuned and gradient- free operation for the separation of empty vs. full capsids. The promise of this system was recognized by 2 market leading rAAV companies – Batavia and Spark – both of which will

support this work by providing hundreds of liters of material, access to their facilities and analytical support. Additional collaborations with MIT and BTEC will assist with process modeling, integration and scale up evaluations. The proposed work will enable: 1) Development of a comprehensive process development toolkit for benchtop

experimental design, modeling and analytics required to execute CCTC capture and polishing operations for rAAV purification 2) Generation of pilot scale process CCTC data with at least 3 rAAV serotypes including at least 2 AAVs from commercial partners Spark and Batavia 3) Integration of CCTC capture and polishing operations for at least 2 rAAV at pilot scale

4) Demonstrated scale up operations in cGMP environment at 50 – 200 L scale 5) Economic modeling via Biosolve platform to enable objective comparison to status-quo platforms and other modalities. We are thrilled to be at the forefront of innovation in such an important industry with a solution that can truly transform clinical and commercial manufacturing of life-saving gene therapies.