Crucial aspects for maintaining rAAV stability

Area of Science:

• Biotechnology
• Molecular Biology
• Gene Therapy

Background:

• Adeno-associated virus (AAV) vectors are critical for gene therapy applications.
• Ensuring consistent product quality and efficacy of AAV vectors during storage is paramount.
• Understanding the impact of storage conditions on AAV vector properties is essential for manufacturing and clinical use.

Purpose of the Study:

• To investigate the influence of various storage conditions on the physicochemical and biological properties of rAAV8 and rAAV9 vectors.
• To assess the short-term storage stability, low-temperature viability, and lyophilization effects on AAV preparations.
• To identify optimal storage strategies for maintaining AAV vector biopotency and efficacy.

Main Methods:

• Quantification of capsid and genome titers.
• Biopotency assessments to evaluate vector functionality and efficacy.
• Analysis of AAV vector stability under different temperature, pH, and storage conditions (short-term, long-term, freeze/thaw, lyophilization).

Main Results:

• Freeze/thaw cycles had minimal impact on rAAV9-aGAL functionality.
• Prolonged room temperature storage decreased biopotency, while elevated temperatures accelerated decay.
• Addition of sucrose and sorbitol enhanced rAAV stability; acidic pH (2.5) significantly reduced biopotency.
• Long-term storage below -65°C preserved rAAV vector effectiveness for years; lyophilization maintained functionality for at least 10 months.

Conclusions:

• Storage temperature and pH significantly impact AAV vector biopotency and efficacy.
• Very low-temperature storage (below -65°C) and lyophilization are effective methods for long-term AAV vector preservation.
• Capsid titer alone does not guarantee preserved biopotency; functional assessments are critical.
• Optimized storage conditions can maximize AAV vector stability and therapeutic potential.