With so many viral-vector-based gene therapies approved by the FDA across different types of viral vectors (AAV, lentivirus, herpes simplex virus 1) and many in late-stage development or clinical trials, the number of approved therapies is growing. There are many challenges that investor-led start-up companies navigate through, from the process of viral-vector manufacturing and CMC analytical method development, to quality issues and regulatory hurdles all in a highly competitive landscape.

This presentation will share experience on tech transfer and CMC challenges for AAV gene therapy products, with a focus on late-phase and preparation for process validation and launch.

In the CGT-sector, plasmids are considered the critical starting material. Within the latest growth in this CGT-sector, the demand for plasmids increased tremendously. Here we present our in-house cutting-edge pDNA production process for off-the-shelf CGT products. We will address our efforts of establishing a robust and scalable process capable of plasmid production for different modalities. By overcoming widespread challenges, we demonstrate production of high-quality plasmids in gram quantity.

Recently, the development of therapies based on recombinant adeno-associated viruses (rAAV) has gained huge interest in the pharmaceutical industry. To ensure a safe and effective gene therapy product, a smart control strategy for the assessment of the rAAVv genome is of utmost importance. In this presentation, different analytical techniques for rAAV genome integrity and identity assessment will be discussed, highlighting their benefits and drawbacks.

Methods for essential biophysical assessments of AAV vectors for in-process sample and drug product at early clinical stages will be introduced. What are the sources of heterogeneity of full particles? Which method among several biophysical techniques is suitable for full and empty ratio determination, depending on sample quality and quantity, and developmental stage?

The development of stable adeno-associated virus (AAV)-based formulations poses many challenges often due to AAV’s intrinsic physico-chemical and structural complexity. Several key vector properties that may impact critical quality attributes of the final product need to be efficiently and reliably characterized and monitored to guide optimal AAV formulation development. Recently, these differences were also noted for AAV drug product of the same serotype but with genome of differing lengths.

This presentation highlights how over a decade of viral vector vaccine development informs gene therapy advancements. Key insights include formulation development, surface interactions, fit-for-purpose analytical methods, modelling, and innovative approaches to create stable, robust gene therapy products.

Purespring Therapeutics utilises a hybrid platform for AAV development in the treatment of kidney diseases, integrating analytical method development at its research facilities with the efficient transfer of technology to manufacturing sites. This strategy ensures rigorous support for product release and stability evaluation. This presentation will highlight case studies of successful method transfers and their application in generating stability data, underscoring advancements in gene therapy for renal disorders.

Downstream processing (DSP) faces challenges due to increased impurities from intensified upstream processing. This presentation will focus on removing host-cell impurities and optimising residual DNA analysis for regulatory compliance. Data on DNA removal for secreted and intracellular AAV serotypes using high-throughput workflows will be presented. DSP strategies for intensified harvest streams, emphasising innovative impurity control and characterisation, will also be discussed.

One of the most significant challenges in adeno-associated virus (AAV) manufacturing is the effective removal of "empty" capsids that lack the gene of interest. Anion exchange chromatography has proven to be the most effective solution for scalable enrichment of full capsids. However, the removal of empty capsids is just one hurdle in this complex process. This work will highlight the critical importance of peak characterisation and its impurity profile, which includes deamidated species, empty and partially filled capsids, host cell DNA, plasmid DNA, host cell proteins, and aggregates.

In the rapidly evolving landscape of mRNA-based vaccines and therapeutics, the demand for robust quality control (QC) measures and advanced analytical capabilities is key. This presentation will discuss our strategy in overcoming QC challenges and strengthening analytical capabilities for mRNA drug substance and lipid nanoparticle product and process development. It will explore traditional QC analytics and emerging analytical trends for deeper product characterisation, ensuring safety, efficacy, and quality. The goal is to share insights, foster open dialogue, and collectively advance the field to support innovation and ultimately benefit global public health.