Abstract
The expanding use of adenoviral vectors for gene therapy has brought about the need for new analytical tools. We have developed an anion-exchange high-performance liquid chromatography method to analyze recombinant adenovirus serotype 5 samples. Before this assay, available analytical methods consisted of either long-term biological assays or required highly purified test articles. These methods were inadequate for optimizing adenovirus production and purification. This assay can quantitate viral particles in either crude lysates or highly pure samples. It can be used to assess particles in both dilute and concentrated samples over a wide dynamic range. Moreover, the population of viral particles eluted in the peak contains most of the infectious virions. This assay is a sensitive technique that overcomes the limitations of previous methods. It provides an essential tool to accomplish process optimization.
Overview summary
We have studied anion-exchange high-performance liquid chromatography as a method to analyze samples containing recombinant type 5 adenovirus. Our results showed that this 20-min assay could be used to quantify viral particles in crude, partially pure, and pure samples. A wide useful range and the ability to handle both dilute and concentrated samples demonstrated that the assay was very robust. This chromatography method could select for a population of particles that contained most of the infectious virions. Because we could apply both crude and pure samples, this assay has become an essential tool for process development.
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