11.21.22 -- AAV Manufacturing: Dual-Plasmid Transfection System Improves Titer & Product Quality

The pursuit of higher rAAV productivity for transient transfection has led to increased focus on the factors that influence the upstream processing. A high cell density process with a novel transfection method has been developed to improve bioreactor vector genome (vg) productivity.

The new system has consistently demonstrated improved productivity and product quality across multiple serotypes, and it can be leveraged to obtain significant process gains in the manufacturing pathways of advanced therapies while maintaining the flexibility of transient transfection.

The market approval of CAR T-cell therapies, together with the continuous emerging number of CAR-based ex vivo and in vivo targeted products, has created huge demand for the supply of large quantities of high-quality-grade lentiviral vectors (LV). An integration of technologies has enabled the successful generation of highly promising suspension PCL LV-CAR clones.

This poster details improvements to the general tbs consensus sequence [KAGNN]x11, enhancements to the sequences within the untranslated region (UTR), and optimization of tbs positioning in relation to the Kozak consensus sequence. The enhanced TRiP technology has been applied to CAR encoding lentiviral vectors, successfully reducing transgene expression in production cells resulting in improved yields.