🔍 They propose that capsid expansion and surface chemistry changes might be key indicators.
💡 Techniques like high-pressure filtration may help separate capsids effectively.
📈 This approach could open new avenues for gene therapy advancements.
Introduction:
The article discusses innovative strategies for separating full and empty adeno-associated virus (AAV) capsids, which is crucial for the production of effective single-gene therapies. Traditional methods, though commonly employed, are often not scalable and depend on serotype, prompting the exploration of structural changes in the AAV capsids as a potential alternative for differentiation.
- Current methods for AAV separation, including ultracentrifugation and anion exchange chromatography, face limitations in scalability and depend heavily on serotype.
- The research suggests that structural changes in AAV capsids associated with encapsulation of DNA may be a viable differentiation approach.
- Four primary structural changes are proposed: capsid expansion due to increased internal pressure with genome packaging, changes in surface chemistry upon DNA interaction, the status of the N-tail structure, and the quantity of the structural protein VP3 on the capsid surface.
- Significant differences in the diameter of empty versus full capsids were noted, with specific attention given to the AAV5 and AAV8 serotypes.
- Potential techniques for separation based on these structural insights include high-pressure methods to deform empty capsids and high-pressure filtration to differentiate based on capsid malleability.
Conclusion:
This research opens new avenues for the separation of full and empty AAV capsids by focusing on structural characteristics, diverging from traditional methods. The findings may lead to more efficient production processes in gene therapies, highlighting the need for further exploration of conformational changes in AAV capsids as potential indicators of their contents.
