Optimization of Tangential Flow Filtration for High-Yield, Scalable Downstream Processing of Adeno-Associated Virus

Area of Science:

• Biotechnology
• Bioprocessing
• Gene Therapy Manufacturing

Background:

• Adeno-associated virus (AAV) production is critical for advancing gene therapies.
• Effective downstream processing, particularly ultrafiltration/diafiltration (UF|DF) using tangential flow filtration (TFF), is essential for high-quality AAV preparations.
• TFF process development requires careful selection of membrane characteristics and optimization of operating parameters to ensure viral functionality and process efficiency.

Purpose of the Study:

• To assess and compare two Sartocon® Hydrosart® TFF cassette architectures (ECO-Screen and E-Screen) for AAV8 clarified lysate UF|DF.
• To define cassette-specific operating regions through flux characterization and small-scale evaluations.
• To provide a quantitative basis for cassette selection and establish a scalable UF|DF framework for AAV purification.

Main Methods:

• Flux characterization and controlled small-scale evaluations of ECO-Screen and E-Screen TFF cassettes.
• Assessment of viral genome retention, host-cell protein, and DNA removal.
• Reproducibility assessments and a 10-fold scale-up study of the UF|DF process.

Main Results:

• Both E-Screen and ECO-Screen cassettes demonstrated high viral genome retention.
• E-Screen achieved faster processing and superior removal of host-cell protein and DNA.
• ECO-Screen enabled efficient operation under reduced pump rates (lower shear), and both formats showed minimal run-to-run variability and predictable scale-up.

Conclusions:

• The study provides a quantitative comparison of TFF cassette performance in AAV purification.
• Both cassette types offer viable options depending on specific process needs (speed vs. shear sensitivity).
• The findings support the rational selection of TFF cassettes and establish a scalable UF|DF framework for AAV manufacturing.