Development of a Novel, Eco-Friendly Mechanical Cell Lysis Process Using Hollow Fiber Filter for AAV Manufacturing

Area of Science:

• Biotechnology
• Gene Therapy Manufacturing
• Cellular Biology

Background:

• Adeno-associated virus (AAV) gene therapy is a promising treatment for genetic disorders.
• Effective recovery of intracellular AAV particles requires efficient cell lysis.
• Existing lysis methods (detergent-based, microfluidization) face challenges in scalability, regulatory compliance, and efficiency.

Purpose of the Study:

• To develop and validate a novel mechanical cell lysis (MCL) method for AAV production.
• To create a scalable, GMP-compatible, and environmentally friendly lysis solution for single-use manufacturing.
• To demonstrate the effectiveness and advantages of MCL over traditional lysis techniques.

Main Methods:

• Development of a novel mechanical cell lysis (MCL) method utilizing hollow fiber filters.
• Implementation in single-use manufacturing systems.
• Parameter optimization and product quality assessment.
• Scale-up validation at 500 L for two cell types.

Main Results:

• Successful proof of concept for the MCL method.
• Demonstrated scalability and compatibility with GMP standards.
• Achieved efficient lysis and AAV recovery across different cell types.
• Validated the method's performance at a 500 L scale.

Conclusions:

• The novel MCL method offers significant advantages over conventional lysis techniques for AAV production.
• MCL provides enhanced versatility, robustness, environmental sustainability, and ease of deployment.
• This method is a superior option for large-scale AAV manufacturing, supporting the advancement of gene therapies.