Executive Summary

Process Analytical Technology (PAT) provides real-time monitoring of defined process parameters (CPPs) and quality attributes (CQAs) during large-scale AAV suspension culture runs. By integrating in-line sensors and analytical tools directly into bioreactors from 50L to 500L+, this approach moves beyond traditional end-point testing to a dynamic, data-driven manufacturing model. The objective is to reduce batch-to-batch variability, increase vector yield and quality, and build a robust data package to support IND submissions within an 18-24 month timeline.

AAV Manufacturing at Scale

This site tour showcases Franklin Biolabs’ preclinical and bioanalytical facilities, spanning over 100,000 sq ft of specialized laboratory and manufacturing-support space.

The Challenge of AAV Process Variability

Scaling adeno-associated virus production from research-grade plasmids to clinical-grade material in large-volume bioreactors introduces significant process variability. Relying solely on terminal batch analysis for CQAs like vector titer, purity, and percentage of full capsids fails to capture the dynamic cellular events that influence these outcomes. A single deviation in metabolic rate or cell health can compromise an entire production run.

This lack of in-process visibility creates downstream risks for chemistry, manufacturing, and controls (CMC) development. It complicates efforts to establish a robust, reproducible process capable of consistently delivering high-quality vectors for IND-enabling toxicology studies and eventual clinical use.

PAT for Real-Time Process Control

Implementing a PAT framework provides the necessary process understanding to mitigate variability in large-scale AAV suspension runs. By integrating probes and automated sampling systems, we can continuously track the key performance indicators of the production culture.

This methodology allows for the creation of a “process signature” for a specific AAV serotype and transgene combination. Deviations from this signature can be identified and corrected in real time. This level of control is fundamental to developing scalable manufacturing strategies accelerating clinical production.

• Live Monitoring: Continuous data streams on VCD, pH, and dissolved oxygen.

• Metabolic Trending: At-line analysis of glucose consumption and lactate production to inform feed strategies.

• Data-Driven Decisions: Adjustments to process parameters are based on quantitative data, not assumptions.

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 provides essential support for our AAV-vector based gene therapy candidate development, and we hope to continue the collaboration for years to come.
— Biotech Partner

Impact on Vector Quality and Regulatory Timelines

A well-controlled manufacturing process is a prerequisite for developing a stable and potent therapeutic product. The ability to produce an AAV-based vector with durable immunogenicity and protective efficacy, as demonstrated in nonhuman primate models (PMID: 34428428), is directly linked to the consistency of the upstream production process. PAT provides the framework to ensure that each batch meets the rigorous quality standards required for clinical evaluation.

This robust process control directly supports an accelerated path to IND. Franklin Biolabs leverages this approach to support its track record, which includes a 100% successful IND rate for programs initiated since 2019 (the Franklin Biolabs brand itself launched in 2024). By minimizing batch failures and generating a comprehensive CMC data package, we help sponsors navigate regulatory requirements and maintain program momentum.