Self-complementary AAV (scAAV) vectors were developed by mutating one of the ITRs to form double-stranded genome. Unlike traditional single-stranded AAVs (ssAAV), scAAVs can enhance gene expression efficiency by bypassing the need for second-strand synthesis. Therefore, it has been widely used in clinical and commercial stages.
In this study, we first presented design improvements in pHelper and pRepCap plasmids that can increase scAAV titers. We also presented a case study for a client-specific Gene of Interest (GOIs), where we apply our Lonza plasmid system. We further improved the scAAV productivity more than 7 fold at the upstream production stage by significantly decreasing transgene plasmid. In summary, we utilized both plasmid engineering and process development strategy to improve self-complementary AAV productivity with clinically relevant GOI.