Proven Intelligence Accelerating Next-Generation Therapies.

Executive Summary

The ratio of full to empty capsids is a primary determinant of AAV vector potency and safety profile. An effective downstream purification process, particularly for prevalent serotypes like AAV8, requires a robust analytical method to resolve these particle populations. This asset details the development of ion-exchange (IEX) chromatography methods for the precise separation and quantification of AAV8 empty and full vector particles, forming the basis for a scalable, GxP-compliant manufacturing process. This approach is foundational to achieving the product quality attributes required for an 18-24 month timeline to IND submission.

The Challenge of AAV Purity

A high concentration of empty capsids in a final AAV vector product presents a known hurdle for clinical translation. These particles can saturate cellular uptake pathways and elicit an immune response without delivering a functional genetic payload. Consequently, regulatory bodies require stringent control over the empty-to-full capsid ratio. The development of a precise, reproducible analytical assay to monitor this quality attribute is the first step in building a compliant manufacturing process.

Our approach is informed by foundational work in developing ion-exchange chromatography assays for the rapid separation and quantification of AAV8 vector particles [PMID: 22428980]. This analytical backbone allows our teams to accurately characterize vector populations and establish clear parameters for downstream purification.

From Analytical Resolution to Scalable Purification

An analytical method, while informative, must be translated into a scalable purification process suitable for large-scale production. The objective shifts from mere quantification to efficient, high-yield recovery of the target full capsids. This requires optimization of column loading, gradient slope, and buffer composition to maximize throughput without compromising purity.

Learn more about the factors for initiating AAV vector programs in the full-length webinar. Watch Now.

Drawing on principles established in scalable tandem column chromatography for viral vectors, we design purification schemes that improve both purity and recovery over legacy methods [PMID: 12427304]. This ensures the final product is stable and meets the specifications for IND-enabling toxicology studies. Strategic Process Blueprints Accelerating Technology Transfer are developed from this data, ensuring a clear path from development to GxP manufacturing.

“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. Franklin Biolabs is a key collaborator in our AAV-vector based gene therapy candidate development and we hope to continue the partnership for years to come.”
— Biotech Partner

Building a Robust CMC Package

Franklin Biolabs provides the process development expertise required to transform a research-grade protocol into an optimized, scalable manufacturing solution. Our work on AAV8 purification is one component of a comprehensive CMC data package designed to meet global regulatory requirements.

• Upstream Process Development: Optimization of transfection parameters and media selection for suspension culture systems from 2L to 500L+.

• Downstream Process Development: Implementation of platform-based or client-specific purification methods, including IEX and affinity chromatography.

• QC Analytics: A suite of over 20 platform assays to characterize titer, purity, potency, and the percentage of full capsids.

This integrated approach, which leverages the continuity of the same scientific team from early development through to technology transfer, supports a 100% successful IND submission rate for programs initiated since 2019, with the Franklin Biolabs brand formally launching in 2024.