(567af) Development of a Single Stage Dual-Ligand Monolithic Chromatography for Plasmid Based Vaccine Production

Many of the production schemes employed by cGMP and GCP facilities combined at least three purification steps to achieve a pharmaceutical grade pDNA vaccine. Increasing the number of unit operations results in product loss and cost ineffectiveness. Although the concept of single-stage pDNA vaccine purification has been found to effectively separate supercoiled pDNA, many problems are still encountered such as low purity, low recovery, high salt consumption, need for post-treatment of pDNA vaccine, and instability of biological-based resin in high affinity separation thus making this process not commercially viable. The present work involves a novel process for pDNA measles vaccine production via optimum fed-batch bacterial fermentation and a single-stage chromatographic purification. Current single-stage chromatography involves only one type of chromatographic mechanism (e.g. anion exchange or affinity interaction) in a one-step loading, washing and elution. The chromatographic technology that is developed in this research is based on a dual-ligand monolithic column that exploits different ligand chemistries in a single column whilst keeping the concept of a single-stage process. The complete details of the invention will be discussed in the full article.