Determination of Full, Partial, and Empty Capsid Ratios for Adeno-Associated Virus (AAV) Analysis

Adeno-associated virus (AAV) is one of the most commonly used delivery vehicles in gene therapy development. However, the presence of product related impurities such as empty and partial AAV capsids co-produced during the manufacturing of AAV vectors could affect the efficacy and safety of AAV products. There are multiple technologies being used concurrently for determining the ratios of these empty or partially filled capsids along with the full AAV. However, these traditional methodologies have their own set of challenges and hence drive a need for an easy to optimize method that, using minute amounts of sample, delivers high resolving power for the determination of empty/full capsid ratios, in about an hour.

 

This webinar demonstrates a robust capillary isoelectric focusing (cIEF)-based method for AAV full and empty capsids analysis. The results show excellent resolution between full and empty capsid as well as potential partial capsid peaks for determination of their ratios. The pI profiles can be determined and used for AAV identification. The results from this methodology correlate well with orthogonal approaches such as anion exchange high performance liquid chromatography (AEX-HPLC). The sample analysis time for this method is rapid, less than 1 hour per sample. The analysis is performed on a well validated and an automated cIEF based platform to obtain reliable and reproducible results across serotypes.

 

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