Abstract
The recent resurgence of using rAAV vectors as commercially viable therapeutic agents has highlighted the need to identify stable capsid/genomic construct combinations which can both endure a typical cGMP manufacturing process and possess a long product shelf life. In this work we investigate the inherent stability of various capsids and genomic constructs using differential scanning calorimetry (DSC). DSC measures the amount of energy required to increase the temperature of a sample and reference and can be used to determine the Tm of the molecule in question. Numerous commercially relevant rAAV serotypes containing several genomic constructs (self-complimentary and oversized genomes) were subjected to melting curve analysis using DSC to compare stability. Additionally, various mobile phase modifiers were added to solution in order to probe the stability of rAAV vector, using organics and kosmotropic/chaotropic salts. These results demonstrate DSC as an effective tool to examine inherent rAAV vector capsid stability and support a role for the serotype and vector genome structure to affect the observed stability.
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