Process Optimization of a DNA Minivector Production System

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The objective of the current project is to collaborate with a current process engineering firm, MemBio to optimize a purification process for production and purification of a high demand DNA minivector generation. The DNA minivectors are linear covalently closed (LCC) molecules known as DNA ministrings or msDNA. Safer DNA vectors with high stability, efficacy and efficient purification are in high demand in the gene/cell therapy industry as a genetic vector for delivery of genetic cargo. Successful completion of this project will generate proprietary processing intellectual property for physically and biochemically optimizing msDNA production and allow them to get it ready for pilot-scale testing. The success of the project will also help strengthen Canadian bio-economy by developing a manufacturing process that can be implemented nationally, and abroad. To date, separation of LCC DNA from other DNA species, both by size and topology has yet to be achieved beyond analytical levels of purification. An efficient and scaled-up approach to generation of mg scale of msDNA and other similar LCC DNA products is needed to drive new developments in genetic medicine. This proposal seeks to implement biochemical, genetic and physical strategies to purify msDNA through the most cost-effective approaches available and generate novel capabilities for MemBio in msDNA and other minivector DNA production.