Accelerating AAV purification process development using high-throughput resin tip module

Area of Science:

• Biotechnology
• Gene Therapy
• Process Development

Background:

• Recombinant adeno-associated viruses (AAVs) are key delivery tools for gene therapy, necessitating robust purification processes.
• AAV purification faces challenges including limited material, high costs, and tight timelines, especially for early-stage development.
• High throughput (HTP) methods offer a solution to overcome these purification development hurdles.

Purpose of the Study:

• To propose and validate an integrated end-to-end HTP workflow for accelerated AAV purification process development.
• To demonstrate the efficiency of HTP methods in determining purification parameters with minimal sample input.
• To assess the impact of the HTP workflow on AAV quality attributes and cost-of-goods.

Main Methods:

• Development of an integrated HTP workflow utilizing a resin tip purification module and an HTP analytical toolkit.
• Screening of purification parameters (binding capacity, resin selection, buffer composition) using micro-volume samples (25 μL resin tip).
• Application of the HTP workflow for AAV quality attribute assessment and comparison with a benchmark method (Robocolumn).

Main Results:

• Efficient determination of purification parameters for separating AAV full, partial, and empty capsids with micro-volume samples.
• Successful prediction of full capsid enrichment and partial capsid removal trends for bench-scale purification.
• Demonstrated comparable AAV quality attributes to benchmark methods, with significantly reduced material volume (2% of benchmark).

Conclusions:

• The integrated HTP workflow effectively accelerates AAV purification process development by leveraging HTP analytical tools for rapid data interpretation.
• This approach significantly reduces material requirements, shortens analytical turnaround times, and offers substantial cost-of-goods savings.
• The validated HTP workflow is crucial for advancing early-stage gene therapy development by enabling efficient AAV purification and quality assessment.