Capillary electrophoresis can be used for a variety of analytical assays to monitor critical quality attributes (CQAs) within the cell and gene therapy field. Plasmids, used as a critical starting material in the manufacture of AAV, require high degree of supercoiling for increased transfection efficiency resulting in increased AAV yields, therefore it is important to monitor plasmid isoform distribution for purity determination. Purity of AAV itself is also a necessary CQA to demonstrate appropriate viral protein expression and expected VP ratio, a key element for correct capsid formation. It is also important to monitor the encapsulated genome to verify that the expected genome size has been packaged. The variety of plasmids required for AAV production can lead to challenging purity analysis based on differences in size and structure. Therefore, the new DNA 20 kb Plasmid and Linear kit from SCIEX was assessed to address this challenge.

Using Pharmaron’s extensive analytical method development experience, here they present an overview of innovative platform capillary gel electrophoresis methods which have been developed to assess AAV and a variety of plasmid sizes.

• What are plasmids and how are they used for viral vector manufacturing
• How can quantitative plasmid purity analysis for a variety of constructs be streamlined
• What information does CGE provide about viral vector quality attributes
• How have developments in technology allowed for high throughput cell and gene therapy analysis