2nd Edition of Cell & Gene Therapy World Conference 2026

Abstract

The AAV-based gene therapy landscape is rapidly expanding beyond rare and ultra-rare diseases to address larger patient populations, creating an increasing demand for scalable, efficient, and cost-effective viral vector manufacturing. However, current AAV production processes continue to face challenges related to productivity, consistency, scalability, and manufacturing costs, contributing to the high cost of gene therapies, which remain among the most expensive on the market. As a critical upstream manufacturing step, transient plasmid DNA transfection plays a key role in determining AAV productivity and process robustness, as inefficiency in DNA delivery can directly impact vector yield and downstream manufacturing. While numerous transfection technologies for AAV production flood the market, limitations remain in achieving robust high-yield performance, maintaining productivity during scale-up, and supporting the flexibility required for modern manufacturing workflows. In this study, we employed a systematic screening and optimization strategy to identify a next-generation transfection approach designed to enhance AAV production performance. The optimized technology was evaluated across multiple AAV serotypes, culture formats, media conditions, and manufacturing scales. Compared with leading commercially available transfection reagents, the optimized approach demonstrated significantly higher vector genome yields while maintaining consistent performance across independent production lots. Importantly, enhanced productivity remained consistent when subjected to diverse upstream process conditions, including variations in culture media and transfection parameters, demonstrating workflow flexibility and robustness. In addition, transfection complex stability studies showed sustained productivity following extended complexation times and low complexation volumes, supporting the operational requirements of large-scale transfection workflows. Scale-up studies further demonstrated that performance established during small-scale development was maintained through a 500-fold increase in culture volume, highlighting the potential for seamless transition from development-scale to commercial-scale manufacturing. Collectively, these findings demonstrate that improvements in transfection technology can address the current limitations of AAV manufacturing by improving vector productivity, process robustness, and supporting scalable production. The ability to consistently achieve high-yield AAV titers across diverse workflows represents an important advancement toward next-generation cell and gene therapies.