Invention: The invention describes modulating the temperature of the solution in an electrospray needle prior to charge-detection mass spectrometry (CD/MS) to enable discrimination between the individual stabilities of multiple forms of viral particles that may be present in an AAV preparation. This increased discrimination provides the operator with a unique ability to determine the stability of the fully assembled viral particles even if the AAV preparation comprises partially filled virions, over-filled virions, and/or various assemblies of capsid proteins lacking a nucleic acid cargo.
Background: Gene therapy and vaccine delivery using adeno-associated virus (AAV) as a vector has emerged as a novel therapeutic modality that has the potential to lead to substantial disease modification in many monogenic disorders, or perhaps even cures. The interest in this approach has been boosted by the recent approval of two AAV-based gene therapies by the United States Food and Drug Administration. Over the last 5 years the number of clinical trials for commercial use have grown tremendously and AAVs are being used more now than ever before. Differential scanning fluorimetry or differential scanning calorimetry are commonly used to measure the thermal stability of AAVs, but these global methods are unable to distinguish the stability of different AAV subpopulations in the same sample.
The goal of this invention is to characterize the stability and unfolding pathways of adeno-associated viral (AAV) capsids, which compose the packaging for efficient viral delivery. Conventional methods for this include charge detection-mass spectrometry (CD-MS) which reports on the solvent exposed surface area of the complex. The method proposed here would be a novel improvement upon current thermal unfolding techniques.
- Gene therapy
- Vaccine development
- Creating model systems
- Medical research and development
- Characterize stability
- Characterize unfolding pathways
- Screen, optimize, and select viral clones