Plasmid Engineering And Process Development To Improve Self-Complementary AAV Productivity

By Chien-Ting Li, R&D Senior Scientist

Self-complementary AAV (scAAV) vectors bypass the need for second-strand synthesis, making them a common choice in clinical and commercial gene therapy programs. However, low productivity remains a challenge for manufacturing economics, creating a need to identify ways to improve output.

Plasmid design offers one potential avenue for optimization. In this study, Lonza Houston’s research and process development teams redesigned the helper and rep/cap plasmids and evaluated their performance against control plasmids across scAAV serotypes 2, 8, and 9. Building on this work, the teams used a client-specific gene of interest in two rounds of design of experiments (DoE) optimization to investigate the effects of different formulations on production performance. Formulations F1 through F13 were assessed by tracking viral genome output and full capsid percentage, providing a basis for comparing productivity and capsid composition across the tested conditions. The optimized approach achieved an increase in upstream productivity of more than sevenfold.

Explore the full dataset to examine viral genome output and full capsid percentages across the formulations and see how plasmid redesign and process optimization influenced the results.