The purpose of this application is to develop a process for improved purification of the non-enveloped viral vector - Adeno-associated virus. This vector is a vehicle for ex vivo and in vivo gene therapy. Starting from crude material, the resulting product must be of high purity and produced in high yields. In addition, this vector must be free from contaminating cellular components such as chromosomal DNA and RNA, protein, membrane components and endotoxins. Requirements for large-scale production of this vector is creating a demand for improved purification processes. Current processes using three (3) consecutive Cesium chloride gradients are cumbersome, inefficient and difficult to scale-up. Alternative technologies based on column chromatography are often inefficient, resulting in low purity and low yields with limited ability to remove contaminants. Column fouling is a particular problem. A new approach involving an innovative technology is proposed. The technology is based on an inert, non-porous, irregularly-shaped polymer using principles of ion-pair reverse-phase chromatography. The unique properties of this resin provide for higher purity and yield and lower residual contaminants. Also, it is reproducible and scalable and will improve economics of vector purification.
The PolyFlo resin is manufactured by Puresyn, Inc. and is to be used as part of the purification process in the development of nucleic acid-based products. The results of the proposed research will offer a process to a new technology area to Academic and Industrial Healthcare Markets. High purity and yield as well as a reduction in process time will lower production expenses from analytical through large-scale development.
