Executive Summary

AAV process intensification focuses on adapting academic-scale protocols for commercially viable manufacturing. The methodology targets scalable suspension culture systems, optimization of plasmid inputs, and refinement of downstream purification to maximize vector yield and purity. These methods are designed to reduce the cost of goods sold (CoGS) for viral vector programs, a factor in accelerating candidates toward IND or Investigational Medicinal Product Dossier (IMPD) submission. Franklin Biolabs executes these CMC strategies within a >100,000 sq ft facility, targeting an 18-24 month timeline to IND.

The Economics of AAV Manufacturing at Scale

The clinical success of AAV-based therapies presents a manufacturing challenge: producing these complex biologicals at a scale and cost that supports broad patient access. Legacy production methods, while sufficient for early discovery, do not provide a viable economic path for indications requiring larger vector quantities. The focus shifts to systematic process intensification to reduce high CoGS, which can otherwise limit the potential of a therapeutic program.

This transition requires a deep understanding of the interplay between vector biology and bioprocess engineering. For example, in programs targeting severe dyslipidemias, the therapeutic concept of in vivo gene replacement is well-established (PMID: 22505953). The task is to build manufacturing processes that can reliably and affordably produce the necessary clinical material for Advanced Therapy Medicinal Product (ATMP) development.

Upstream Optimization: From Flasks to Bioreactors

A foundational step in process intensification is the transition from adherent cell culture to suspension-based systems. While adherent platforms are suitable for initial vector production, they present significant scalability limitations.

• Suspension Culture: Utilizing single-use bioreactors (2L to 500L+) for suspension culture provides a direct, scalable path. This approach allows for precise control over environmental parameters like pH, dissolved oxygen, and nutrient feeding, leading to higher cell densities and improved volumetric productivity.

• Plasmid & Transfection: The efficiency of the triple transfection process is a major determinant of final vector yield. Process development focuses on optimizing plasmid DNA quality, purity, and the stoichiometric ratios of the rep/cap, helper, and gene-of-interest plasmids.

• Vector Design Considerations: The intrinsic producibility of a vector is also a factor. A well-designed construct, such as a muscle-tropic AAV9 vector engineered for high expression, can provide a significant advantage before manufacturing even begins, simplifying the path to rescuing a disease phenotype (PMID: 34454844).

“We started collaborating with UPenn Vector core in 2023 and the AAV vector which they manufactured laid a foundation for development of a gene therapy candidate which will enter soon preclinical studies. The key people from UPenn Vector Core joined Franklin Biolabs and our partnership transitioned without interruption from UPenn Vecor Core to Franklin Biolabs Research Vector Division. The collaboration with Franklin Biolabs is integral to our AAV-vector based gene therapy candidate development, and we look forward to continuing the partnership for years to come.”