Separating Empty and Full Recombinant Adeno-Associated Virus Particles Using Isocratic Anion Exchange Chromatography.

Area of Science:

• Biotechnology
• Gene Therapy
• Chromatography

Background:

• Recombinant adeno-associated virus (rAAV) gene therapies are critical biotherapeutics.
• Production challenges include the presence of empty AAV particles, impacting safety and efficacy.
• Efficient purification methods are needed to separate empty and full rAAV particles at scale.

Purpose of the Study:

• To develop a robust and scalable purification process for separating empty and full rAAV particles.
• To present a novel anion exchange chromatography (AEC) method for enriching full rAAV2 particles.

Main Methods:

• Development of a novel anion exchange chromatography process utilizing isocratic wash and elution steps.
• Identification of an operating design space to ensure process robustness.
• Comparison of isocratic chromatography with traditional shallow linear gradient elution.

Main Results:

• The developed isocratic AEC process effectively enriches full rAAV2 particles.
• The process demonstrates robustness within the identified operating design space.
• Isocratic chromatography offers advantages over gradient elution, including reduced buffer consumption and smaller pool volumes.

Conclusions:

• A novel and robust isocratic anion exchange chromatography process has been established for purifying full rAAV particles.
• This method addresses the challenge of separating empty and full AAV particles in large-scale manufacturing.
• The isocratic approach enhances manufacturing efficiency and robustness for rAAV gene therapies.