Novel, Targeted Method for Bacteriophage Purification

PROJECT SUMMARY The fields of gene therapy and bacteriophage therapy are rapidly expanding and show promising potential for treating genetic disorders and antibiotic resistant bacterial infections, respectively. To meet these needs, improved high throughput and high affinity viral purification methods will be required. Current high throughput viral purification methods mainly consist of resin-based chromatography coupled with multiple filtration steps. Chromatography-based methods often lack specificity for the particular virus target. For example, the commonly used ion exchange resins are not able to separate contaminants sharing similar surface net charges to the virus particles. Selection of mixed virus varieties such as viruses of different serotypes cannot be achieved with chromatography methods. Affinity-based resin chromatography products targeting viral vectors exist but are expensive or not suitable for large-scale purification. Additionally, they involve coupling a binding entity to resin, which presents several disadvantages, including leaching of the affinity ligand from the chromatography support or the ligand co-eluting with the virus particles. Thus, improved methods for simple, rapid, and scalable purification of viral vectors are needed to serve both the small-scale research community and large-scale industrial production. Bondwell Technologies proposes to develop a novel purification system for large biomolecules, such as viral vectors. The unique qualities of our functionalized biomaterial provide tremendous versatility and addresses the current limitations of small- and large-scale viral purification. During this proposed Phase I effort, we will first functionalize our biomaterials with an entity capable of specifically recognizing a viral target. We will then test the biomaterial for their ability to bind the viral target. Finally, we will prepare our biomaterial to be used as a membrane and test its ability to bind a viral target with different complexities of starting material (e.g., with cellular debris present).