Practical Methods for Supercoiled pDNA Production

Increased demand for plasmid DNA (pDNA) to be produced to tighter and more exacting specifications, even for early preclinical work, has led to many researchers and manufacturers reevaluating their production methodologies. This chapter is intended to offer realistic methods that may be employed by those wishing to purify between 100 and 200 mg of pDNA in-house based on availability of equipment and other resources. This scale of production typically requires a compromise between techniques used with gravity-flow or vacuum devices and intermediate scale column chromatography. The methodologies described in most instances are unit processes that can be adapted into an appropriate scheme given such considerations as the desired purity level or the quality of the feedstock.