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In Vitro Disassembly of Influenza A Virus Capsids by Gradient Centrifugation
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DNA Released by Adeno-Associated Virus Strongly Alters Capsid Aggregation Kinetics in a Physiological Solution
Curtis W Jarand1, Karen Baker2, Matthew Petroff2
1Department of Physics, Tulane University, New Orleans, Louisiana 70118, United States.
Biomacromolecules
|April 29, 2024
Summary
Adeno-associated virus (AAV) stability is crucial for gene therapy. This study reveals how AAV capsid interactions with DNA influence degradation and aggregation in physiological conditions.
Area of Science:
- Biotechnology and Biomedical Engineering
- Molecular and Cellular Biology
- Virology
Background:
- Adeno-associated virus (AAV) is a key vector in gene therapy, but its stability and degradation pathways are not fully understood.
- Understanding AAV behavior under physiological conditions is essential for optimizing gene therapy applications.
Purpose of the Study:
- To investigate the degradation and aggregation mechanisms of an engineered AAV serotype under physiological pH and ionic strength.
- To elucidate the role of encapsulated DNA and capsid interactions in AAV stability.
Main Methods:
- Incubation of AAV particles with varying DNA fractions at temperatures ranging from 30 to 60 °C.
- Measurement of changes in molecular weight over time using total light scattering intensity at 90°.
Main Results:
- Full AAV vectors showed rapid DNA release and subsequent slow aggregation.
- Empty AAV vectors exhibited immediate, rapid colloid-type aggregation.
- Mixtures of full and empty capsids displayed reduced initial aggregation, suggesting interactions between capsids and released DNA.
Conclusions:
- AAV capsid-DNA interactions significantly influence degradation, aggregation, and DNA release kinetics in physiological solutions.
- These findings provide critical insights into AAV vector stability for gene therapy development.
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