Rapid growth in the cell and gene therapy industry has generated an urgent need for robust analytics around characterization of both vectors and nucleic acid biotherapeutics such as oligonucleotides, plasmid DNA, small RNA, mRNA and double stranded DNA.

 

Adeno-associated virus (AAV) is one of the most widely used gene delivery vehicles for gene therapy because of its non-pathogenicity, low immunogenicity and different tropism to multiple cell types. Although there are efforts to develop methods for characterization of AAV, there are still limitations and drawbacks to some of these analyses. In addition, the different workflows are done on different platforms. Here we propose multiple key analytical workflows that can be performed on a single Capillary electrophoresis (CE) based platform for extensive characterization of AAV. Some of these methodologies include AAV capsid purity analysis (VP1, VP2 and VP3) using both UV and laser-induced fluorescence (LIF) detection.

 

In addition, a suite of analytical methods recently developed and optimized for characterization of the nucleic acid biotherapeutics and detection of impurities. Presented here is a ready-to-use protocol for not only in-depth characterization of various nucleic acid but also analysis of impurities in these biotherapeutics. Furthermore, all of these methods for vector and nucleic acid analysis are developed on a GMP compliant instrument with associated reagent kits and consumables commercially available; thus, making it a seamless process to analyze these newer modalities.

 

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