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

Qingxuan Li1, Mahsa Hadidi1, Steven Benner2

  • 1Sanofi Genomic Medicine Unit, CMC, Purification Process Development, Waltham, Massachusetts, USA.

Biotechnology Progress
|July 18, 2025
PubMed
Summary
This summary is machine-generated.

Developing high throughput (HTP) methods accelerates adeno-associated virus (AAV) purification. An integrated HTP workflow using resin tips and analytical tools significantly reduces material requirements and costs for gene therapy development.

Keywords:
AAV purificationHTP workflowanalytical toolkitgene therapyresin tip

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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.