Platform for Rapid Expression, Purification, and Analysis of Patient-Specific Gene Therapy Products

点击翻译按钮获取中文摘要

ABSTRCT By 2050, adeno-associated viruses (AAVs) will be established as the workhorse for in vivo gene therapy, impacting 60 - 80 million patients per year worldwide, and the success of this gene therapy will rely on the ability to manufacture application-/patient-specific AAVs affordably and rapidly. However, while in vivo and ex vivo gene therapy products are advancing rapidly, their biomanufacturing technology has not seen comparable progress. At present, manufacturers achieve the mandated product purity upon strenuous optimization - at the expense of productivity and product efficacy and run multiple production batches to meet the clinical demand -with significant delays and cost to patients (up to $1.2M per individual). To address the challenges faced by current AAV biomanufacturing, we propose to develop an integrated purification and sensing technologies to accelerate process design and optimization and make small-batch AAV production affordable to specific patient cohorts. The technology is based on the integration of chromatographic adsorbents for isolation of “full” AAVs in flow- through mode coupled with label-free impedimetric biosensors for quantifying AAV titer and full:empty ratio. These technologies will enable the application of advanced analytics for the prediction of AAV therapeutic efficacy by correlating process parameters with biomolecular-level CQAs of AAVs. The integrated purification and biosensor technology will be demonstrated in a pilot process for producing patient-specific doses of rAAV2- retro and scAAV9-CB-CLN6, which have been shown to correct brain and behavioral pathologies in CLN6- Batten disease, a rare lysosomal storage disease.