Recombinant adeno-associated virus (rAAV) is a small (25-nm) virus that is composed of a protein shell called a capsid and a single-stranded DNA genome containing the transgene and other structural and regulatory elements. During the process of packaging the transgene into the rAAV capsid, errors can happen leading to packaging of truncated genome as well as fragments from host cell genome and plasmid vector. Size determination of rAAV genome offers insights in assessing the transgene integrity of the rAAV.

 

Verification of rAAV genome size has been traditionally done by denaturing agarose-gel electrophoresis and southern blot. Both are time consuming and with limited precision and resolution on size determination. In this webinar, we present a genome sizing methodology for assessing AAV transgene integrity using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). Our results indicate that this method is fast and robust, capable of separating intact and partial AAV genome as well as small size impurities with excellent resolution and repeatability.

 

 

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