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Development of a novel purification method for AAV vectors using tangential flow filtration
Rimi Miyaoka1,2, Yuji Tsunekawa1, Yae Kurosawa1,3
1Division of Molecular and Medical Genetics, Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
Biotechnology and Bioengineering
|August 16, 2023
Summary
This study introduces a novel surfactant-enhanced tangential flow filtration (TFF) method for purifying adeno-associated virus (AAV) vectors. This efficient process significantly reduces protein contaminants, enabling high-quality gene therapy production.
Area of Science:
- Biotechnology
- Gene Therapy
- Virology
Background:
- Adeno-associated virus (AAV) vectors are crucial for gene therapy, but complex manufacturing leads to high costs.
- Current purification methods for AAV vectors are often intricate and contribute to elevated product pricing.
Purpose of the Study:
- To develop a simplified and efficient high-quality purification method for adeno-associated virus (AAV) vectors.
- To reduce manufacturing costs associated with AAV vector-based gene therapies.
Main Methods:
- Utilized tangential flow filtration (TFF) combined with anionic and zwitterionic surfactants.
- Developed a novel purification strategy to inhibit protein aggregation during AAV vector processing.
Main Results:
- Achieved 99.5% clearance of residual proteins from the crude AAV vector product.
- Confirmed the infectivity of the purified AAV vector in vitro and in vivo.
- Observed no significant inflammation or tissue damage in mouse skeletal muscle post-administration.
Conclusions:
- The novel surfactant-assisted TFF method provides an efficient platform for high-quality AAV vector purification.
- This approach has the potential to lower the cost of AAV vector-based gene therapies.
- The method ensures the safety and efficacy of purified AAV vectors for therapeutic applications.
