Successfully advancing a gene therapy program requires effectively combining internal capabilities with external partners. This presentation provides an overview of AAVantgarde Bio’s approach to positioning a gene therapy program for success through strategic use of CDMOs and focused internal expertise. Topics include the rationale and framework used for CDMO selection, alignment of development and manufacturing strategies, and integration of process development and analytical data to support decision-making. The talk highlights how coordinated execution across internal teams and external partners can accelerate progress, manage complexity, and establish a strong foundation for clinical advancement.

Bringing a gene therapy to clinic presents unique CMC challenges requiring strategic foresight and coordination. This presentation shares real-world lessons in preparing CMC packages for clinical trials, focusing on early-phase development, regulatory readiness, and product comparability and justification of specifications. It highlights strategies for robust documentation, managing process changes, and logistics affecting manufacturing timelines. Strategic planning and foresight are essential to overcoming CMC hurdles and enabling smooth clinical progression.

Recombinant adeno-associated virus (rAAV) vectors are pivotal for gene therapy, but their manufacturing remains expensive, impacting the overall product development costs and ultimately limiting accessibility for patients. This presentation explores the hurdles and technical avenues for enhancing rAAV process productivity with the aim of reducing the cost per viral genome through cutting-edge bioprocessing technologies.

The Cell and Gene Therapy Catapult (CGTC) is advancing cost-effective, next-generation rAAV manufacturing by developing continuous and intensified upstream processes that enhance productivity, scalability, and consistency. Complementing this, process and mechanistic modelling, integrated with Process Analytical Technology (PAT) and automation, provides real-time monitoring, predictive insights, and adaptive control. Together, these innovations create smart-by-design platforms that accelerate development, reduce costs, and enable robust rAAV manufacturing at commercial scale.

Adeno-associated virus (AAV) vectors represent promising gene therapy tools, but manufacturing challenges including poor secretion and suboptimal empty-to-full capsid ratios, limit their therapeutic potential. This presentation describes comprehensive engineering biology approaches developed by our research group to address these limitations. We systematically optimised transient transfection conditions, engineered novel capsid variants with improved properties, and modified the membrane-associated accessory protein (MAAP), to enhance vector secretion and packaging efficiency. Our integrated engineering strategies demonstrate how rational design principles can overcome key bottlenecks in AAV production, offering pathways toward more efficient and scalable manufacturing processes for next-generation gene therapies.

This presentation examines the unique manufacturing complexities of dual AAV vector systems including vector design, co-packaging efficiency, and ensuring balanced expression of both halves. It highlights key control strategies for process optimisation, analytical characterisation, and product consistency. Emphasis is placed on developing robust, scalable workflows to maintain quality and regulatory compliance in dual-vector gene therapy manufacturing.

Adeno-associated virus (AAV) vectors represent one of the most versatile clinical platforms for gene delivery. However, AAV vectors transduce cells poorly, resulting in the administration of exceedingly high doses in the clinic, which increases the risk of adverse events. To improve vector cargo delivery capacity and transduction efficiency, we are rationally designing novel capsids. We assessed and identified different sites for capsid engineering that do not impact vector yields while enhancing transduction.

For scale-up of virus formulation processing, we have developed a scale-down freezing system that mimics large-scale freezing and thawing. This system serves dual purposes: it acts as a stress model for formulation development, aiding the selection of excipients. Furthermore, it provides insights into the microscopic and macroscopic effects expected in large scale operations, such as ice front formation, cryoconcentration, and phase separation after thaw.

CRISPR technologies enable precise correction of disease-causing mutations but require transient expression to minimise off-target effects. Delivering ribonucleoproteins (RNPs) via virus-like particles (VLPs) to target cells, provides a safer alternative. In proof-of-concept experiments, VLPs pseudotyped with various viral glycoproteins targeted lung cells carrying a mutated GFP gene. Our results demonstrated the successful delivery of editing components, precise correction of the mutation, and restoration of fluorescence, thereby emphasising VLPs’ therapeutic potential.

RedTail is a gene therapy platform using enveloped vaccinia virus for systemic, tumor-targeted delivery. It resists immune clearance and enables direct payload delivery to metastatic sites. CLD-401 expresses an IL-15 superagonist and evades complement lysis and antibodies. Manufacturing of the enveloped RedTail products requires specialised processes to maintain membrane integrity and ensure consistent potency.

This presentation highlights analytical approaches used to evaluate the efficacy and quality of AAV-based gene therapies. It discusses general strategies for characterising vector components, assessing potency, and ensuring product consistency. Emphasis is placed on the importance of reliable analytical tools to support development, manufacturing, and regulatory expectations across the gene therapy lifecycle.

Non-wild type AAVs are engineered AAVs which improve the targeting efficiency, safety, and specificity for therapeutic applications. The behavior of modified AAVs in CE-based techniques was investigated using labchip, CE-UV, and other techniques. The study revealed the presence of temperature-dependent aggregate peaks that failed to migrate according to their expected size. These anomalous migration patterns were observed across varying conditions, suggesting the resulting electropherogram contained artifact peaks.