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Design of experiments as a decision tool for cell therapy manufacturing
Esmond Lee1, Devin Shah2, Matthew Porteus3
1Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Cytotherapy
|March 1, 2022
Summary
Optimizing gene editing for cell therapies using design of experiments (DoE) identified cost-effective parameters. This approach reduces manufacturing costs for advanced therapeutics like those for IPEX syndrome.
Area of Science:
- Biotechnology
- Gene Therapy
- Process Development
Background:
- Cell therapies are expensive due to complex manufacturing and autologous production.
- Reducing manufacturing costs is crucial for broader cell therapy accessibility.
Purpose of the Study:
- To apply a design of experiments (DoE) approach to optimize the gene editing process for a cell therapy.
- To identify key cost drivers in cell therapy manufacturing.
Main Methods:
- Utilized DoE to analyze factors influencing CRISPR/Cas9 gene editing efficiency in CD4+ T lymphocytes.
- Investigated the impact of AAV multiplicity of infection (MOI) and sgRNA amount on gene editing.
- Performed cost analysis across the design space to determine optimal, cost-effective parameters.
Main Results:
- AAV MOI and sgRNA amount were significant factors affecting gene editing efficiency.
- Highest MOI and sgRNA yielded the best editing frequency.
- Optimal cost-effective gene editing occurred at 193,000 vg/cell AAV and 1.78 μg sgRNA.
Conclusions:
- DoE provides a data-driven, efficient method for understanding and optimizing complex biological manufacturing processes.
- This approach can lead to more robust and cost-effective strategies for cell therapy production.
- The findings support the development of investigational therapeutics for conditions like IPEX syndrome.
