Insight Into the Degradation Pathways of an AAV9.

Area of Science:

• Biotechnology
• Gene Therapy
• Molecular Biology

Background:

• Recombinant adeno-associated virus (AAV) vectors are crucial for in vivo gene therapy, with numerous clinical trials underway.
• Limited studies exist on AAV degradation pathways due to material constraints and structural complexity.
• Understanding AAV stability is vital for successful gene therapy applications.

Purpose of the Study:

• To elucidate the degradation pathways of a model AAV9 vector.
• To assess the impact of various stress conditions on AAV9 critical quality attributes and transgene expression.
• To identify risks associated with AAV9 formulation and storage.

Main Methods:

• A model AAV9 vector with a GFP transgene was subjected to various stress conditions.
• Critical quality attributes of the AAV9 vector were assessed.
• Transgene expression levels were monitored under different stress conditions.

Main Results:

• AAV9 exhibited degradation under thermal stress at 25°C and 40°C.
• Base formulation posed a risk for AAV9 instability and potency loss under thermal stress.
• AAV9 remained stable under freeze-thaw, interfacial stress (membrane filtration), and short-term cold storage (4 weeks at 5°C).

Conclusions:

• Thermal stress significantly impacts AAV9 stability and potency.
• Formulation strategies are critical to mitigate AAV9 degradation under specific stress conditions.
• AAV9 demonstrates robustness under freeze-thaw, filtration, and short-term cold storage, suggesting suitability for certain handling conditions.