Manufacturing Process Intensification of Adeno-Associated Viral Vectors Type-8 Using Weak Partitioning Chromatography...

Area of Science:

• Biotechnology
• Bioprocessing
• Gene Therapy Manufacturing

Background:

• Recombinant adeno-associated viral vector (AAV) production requires efficient separation of full and empty capsids for gene therapies.
• Current anion exchange chromatography (AEX) methods face scalability limits due to low loading capacities (2e13-2e14 capsids/mL) with increasing upstream titers.
• Monoliths and membranes are preferred for AAV purification over resins to avoid diffusion limitations.

Purpose of the Study:

• To develop a next-generation purification process for AAV with significantly higher loading capacity (>1e15 capsids/mL).
• To improve the scalability, robustness, and efficiency of full/empty capsid separation in AAV manufacturing.
• To address the limitations of current linear gradient AEX methods in high-titer AAV production.

Main Methods:

• A scalable weak partitioning anion exchange chromatography (AEX) method using isocratic elution was developed.
• The method was demonstrated on CIM QA monoliths, commonly used in AAV manufacturing.
• Process optimization included load conditions and UV-based signal automation.

Main Results:

• The weak partitioning AEX method achieved over 80% full capsid purity and over 80% genomic yield.
• This approach enables over 10-fold higher loading capacity per cycle compared to standard linear gradient AEX.
• Processing time was reduced by 10-fold, facilitating rapid cycling or continuous processing.

Conclusions:

• Weak partitioning AEX with isocratic elution offers a scalable and efficient solution for high-titer AAV purification.
• The method overcomes the limitations of linear gradient AEX, providing higher purity and yield.
• This next-generation process supports the demands of advanced gene therapy manufacturing.