An Integrated, Scalable Platform for rAAV Vector Development Designed for Translational Success

Thursday, March 14, 2024

11:00 AM-1:00 PM

BIOMED PhD Thesis Defense

Title:
An Integrated, Scalable Platform for rAAV Vector Development Designed for Translational Success

Speaker:
Xueyuan Liu, PhD Candidate
School of Biomedical Engineering, Science and Health Systems
Drexel University

Advisors:
Beverly Davidson, PhD
Professor of Pathology and Laboratory Medicine
Director, Raymond G. Perelman Center for Cellular and Molecular Therapeutics
Perelman School of Medicine
University of Pennsylvania
Chief Scientific Strategy Officer (CSS)
Children's Hospital of Philadelphia (CHOP) Research Institute

Lin Han, PhD
Professor
School of Biomedical Engineering, Science and Health Systems
Drexel University

Details:
Recombinant adeno-associated virus (rAAV) is a leading viral vector modality for gene therapy, with several approved treatments and hundreds of ongoing clinical trials. rAAV manufacturing, while sufficient to support preclinic studies and clinical trials, is challenged at scale-up due to the translational gap between vector development and manufacturing. Traditionally, vector development is limited to the early stage, relying on basic purification processes and analytical tools. It generally focuses on expression, target specificity, and immunogenicity, neglecting the crucial aspects of vector manufacturability: productivity and purity, which are vital for the clinical success of novel therapeutic vectors.

I hypothesized that early identification of vector manufacturability is critical and necessary to assist with the translation of novel vectors. To address this, I proposed creating an integrated, scalable, high throughput rAAV screening and optimization platform to overcome vector development and manufacturing challenges at the discovery and pre-clinic stages.

This research proceeded with three aims: (1) Develop a microscale rAAV production method for high throughput productivity screening, enabling simultaneous testing of multiple constructs and conditions at a reduced cost and time without needing special equipment and expertise. It also provides a viable option to optimize vector design, modulate the effects of packaging plasmids, and maximize production efficiency with a design of experiments (DOE) approach. The platform is scalable, reflecting productivity propensity at scale. (2) Validate and integrate a novel UV-VIS/DLS/SLS analytic platform and additional methods like CsCl density banding, capillary electrophoresis, Nanopore sequencing, and multiplex ddPCR to screen vector purity. The platform, capable of quantifying and characterizing rAAV vectors in high throughput, eliminates the need for reference standards, large number of purified vectors, and intensive training and is suitable for vector development at the early discovery stage. The combinatorial analytic platform identifies product-related impurities and optimizes vector purity early on. (3) Demonstrate the utility of the integrated platform via developing a novel FVIII vector for Hemophilia A. In conjunction with in vivo testing, the integrated platform ensures optimized vector manufacturability and efficacies for clinic success.

In summary, the integrated, scalable screening platform bridges the gap between discovery and clinical manufacturing, enhancing vector manufacturability throughout therapeutic development. This improvement moves novel therapies closer and faster to patients, highlighting the importance of early manufacturability assessment in the successful gene therapy development.

Contact Information

Natalia Broz
njb33@drexel.edu