Presentation Information
[P04-532]Development of a Novel Separation Membrane to Reduce Fouling in the AAV Vector Clarification Process
○Kota Tsukamoto1, Hirokazu Sakaguchi1, Taisuke Kogawa1, Takeo Teranishi1 (1. Toray Industries, Inc (Japan))
Keywords:
filtration membrane,AAV,clarification,Gene therapy,Biopharmaceuticals,fouling
Biopharmaceuticals are attracting increasing attention due to their higher therapeutic efficacy compared with conventional small-molecule drugs, and the global market continues to expand. In recent years, gene therapy, which treats diseases caused by genetic defects by delivering functional genes, has emerged as a promising therapeutic approach. Viral vectors are commonly used as gene delivery vehicles, and among them, adeno-associated virus (AAV) has been widely developed because of its low pathogenicity, low immunogenicity, and low risk of host genome integration.
However, AAV manufacturing processes are still under development. Because cell lysis is performed after cell culture, AAV harvests contain large amounts of impurities, which often cause membrane fouling and clogging during the clarification step. This clarification step is a critical process connecting cell culture and downstream purification, and it requires efficient recovery of AAV while achieving sufficient clarity to prevent clogging in subsequent chromatographic steps.
To address these challenges, we developed a novel clarification membrane for AAV processing by integrating separation membrane technology with a low-fouling modification. A nonwoven-based membrane with low pressure drop and high particle capture performance was applied to the first-stage coarse filtration, while a hollow fiber ultrafiltration membrane, developed based on hemodialysis membrane technology, was used for the second-stage filtration. In addition, a proprietary polymer was introduced to hydrophilize the membrane and significantly suppress the adhesion of biological components, resulting in a membrane structure and design that is highly resistant to clogging even for impurity-rich AAV harvests.
The performance of the coarse filtration membrane was evaluated by relative comparison with benchmark commercial products under identical processed volume per membrane area. The developed membrane successfully removed impurities in the range of 3–10 µm that could not be sufficiently removed by the benchmark products, and a significant reduction in turbidity was observed. Furthermore, the filtration pressure at a given processed volume was reduced, indicating improved clarification efficiency and extended membrane lifetime.When evaluated as an integrated clarification module combining the coarse filtration and ultrafiltration membranes, the developed system achieved more than four times higher filtration capacity compared with benchmark modules, demonstrating markedly improved resistance to clogging. The filtrate met the required turbidity criteria and showed no issues in process compatibility with subsequent affinity purification steps.
These results demonstrate that the developed clarification membrane can simultaneously achieve high clarity, reduced fouling, and excellent process compatibility in AAV manufacturing. Application of this membrane to AAV production processes is expected to contribute to improved productivity, consistent product quality, and reduced manufacturing costs.
However, AAV manufacturing processes are still under development. Because cell lysis is performed after cell culture, AAV harvests contain large amounts of impurities, which often cause membrane fouling and clogging during the clarification step. This clarification step is a critical process connecting cell culture and downstream purification, and it requires efficient recovery of AAV while achieving sufficient clarity to prevent clogging in subsequent chromatographic steps.
To address these challenges, we developed a novel clarification membrane for AAV processing by integrating separation membrane technology with a low-fouling modification. A nonwoven-based membrane with low pressure drop and high particle capture performance was applied to the first-stage coarse filtration, while a hollow fiber ultrafiltration membrane, developed based on hemodialysis membrane technology, was used for the second-stage filtration. In addition, a proprietary polymer was introduced to hydrophilize the membrane and significantly suppress the adhesion of biological components, resulting in a membrane structure and design that is highly resistant to clogging even for impurity-rich AAV harvests.
The performance of the coarse filtration membrane was evaluated by relative comparison with benchmark commercial products under identical processed volume per membrane area. The developed membrane successfully removed impurities in the range of 3–10 µm that could not be sufficiently removed by the benchmark products, and a significant reduction in turbidity was observed. Furthermore, the filtration pressure at a given processed volume was reduced, indicating improved clarification efficiency and extended membrane lifetime.When evaluated as an integrated clarification module combining the coarse filtration and ultrafiltration membranes, the developed system achieved more than four times higher filtration capacity compared with benchmark modules, demonstrating markedly improved resistance to clogging. The filtrate met the required turbidity criteria and showed no issues in process compatibility with subsequent affinity purification steps.
These results demonstrate that the developed clarification membrane can simultaneously achieve high clarity, reduced fouling, and excellent process compatibility in AAV manufacturing. Application of this membrane to AAV production processes is expected to contribute to improved productivity, consistent product quality, and reduced manufacturing costs.
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