Expanding AAV Process and Characterization Analytical Technologies

Gene therapies promise to become revolutionary curative treatments, yet realization of this potential requires advances in biomolecular understanding to be on par with quality standards of today’s state-of-the-art treatments (e.g., monoclonal antibodies).
Categories
Cell and Gene therapies
Assays
Project status
100% Completed

Industry Need

  • Gene therapies promise to become revolutionary curative treatments, yet realization of this potential requires advances in biomolecular understanding to be on par with quality standards of today’s state-of-the-art treatments (e.g., monoclonal antibodies). 
  • Characterization of mAbs has evolved during the past 30 years to include an advanced toolbox of measurements to clarify identity, quality, and stability. In turn, numerous structure/function and structure/stability correlations have been identified, allowing product quality attribute (PQA) criticality to be assessed using a risk-based approach. 
  • Innovation and adaptation beyond existing high-resolution methods are needed to build similar knowledge around gene therapy delivery vectors. 


Solution

NC State, in partnership with NIST and Pfizer, aims to pioneer the combination of multi-attribute method (MAM) mass spectrometry and ion mobility spectrometry (IMS) into the gene therapy space. Modality-targeted development will provide the specificity and speed necessary to meet the emerging demands of process development and set the industry standard for product knowledge and control. 

Outputs/Deliverables

  • Generated an industry-grade AAV2 suitable for analytical method innovation
  • MAM-based attribute-specific control for modification susceptible peptides (Target <15% CV or limit specification)
  • MAM-based purity test with limit of detection (Target < 0.5 % Impurity)
  • Develop first-of-its-kind IMS VP subunit assay (Target throughput <15 min, complete VP resolution, serotype identity)
  • Enable quality-by-design of future AAV materials via an associated transduction assay (repeatability target < 30 and intermediate precision target < 40% CV)

Impacts

By combining MAM and IMS in gene therapy, this project will enable increased speed, resolution, and robustness of analytical and biological methods to characterize gene therapy products, which can lead to better control of product quality.

Publications

Ryan, J. P., Kostelic, M. M., Hsieh, C., Powers, J., Aspinwall, C., Dodds, J. N., Schiel, J. E., Marty, M. T., & Baker, E. S. (2023). Characterizing Adeno-Associated Virus Capsids with Both Denaturing and Intact Analysis Methods. Journal of the American Society for Mass Spectrometry, 34(12), 2811-2821. https://doi.org/10.1021/jasms.3c00321

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Project Lead

North Carolina State University

North Carolina State University

Participating Organizations

Federal Stakeholder: National Institute of Standards and Technology

Federal Stakeholder: National Institute of Standards and Technology

Pfizer, Inc.

Pfizer, Inc.