SBIR-STTR Award

The purpose of this application is to develop commercial processes for improved purification of non-enveloped viral vectors such as adenovirus. These vectors are vehicles for vaccination or 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, these vectors must be free from contaminating cellular components such as chromosomal DNA and RNA, protein, membrane components and endotoxins. Requirements for large scale production of these vectors is creating a demand for improved purification processes. Current processes using 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 yields and limited abilities 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 improved purity and yield resulting improved economics for vector purification.

Puresyn, Inc. has developed a purification process for recombinant adenovirus vectors utilizing its proprietary chromatographic resin, PolyFlo(R). In Phase I we demonstrated that a tandem process of an ion- exchange and PolyFlo chromatography results in product of high purity and yield that exceeds cesium chloride. In addition, we demonstrated that PolyFlo has a high capacity for host and viral contaminant removal and the process is reproducible and scaleable. We are seeking Phase II funding to concentrate on recombinant adenovirus purification to include large-scale purification up to 10(16) particles per run without compromising purity, yield or biological activity. We will refine and expand our current chromatography protocol and evaluate the total process, lysis through formulation. In addition, we will conduct a mouse study to determine the correlation between purity, biological activity and safety. We propose to evaluate the effects of purity levels on immune and toxic responses to the vector. Results of this research will have immediate impact for those engaged in the use of recombinant vectors for gene delivery and have commercialization implications for PolyFlo. Additionally, the ability to purify at large-scale will eliminate a significant obstacle to the development of the Gene Therapy Market. PROPOSED COMMERCIAL APPLICATIONS: The PolyFlo(R) resin is manufactured by Puresyn, Inc. and is being used as part of the purification process in the development of nucleic acid based products. We anticipate that the Phase II aims will be achieved and support the marketing of PolyFlo to the Academic and Industrial Healthcare Market for the purification of rAd. Additionally, the ability to purify at large-scale will eliminate a significant obstacle to the development of the Gene Therapy Market.

Public Health Relevance:
This Public Health Relevance is not available.

Thesaurus Terms:
Adenoviridae, Ion Exchange Chromatography, Recombinant Virus, Resin, Technology /Technique Development, Transfection Vector Gene Therapy, Protein Purification Immunochemistry, Laboratory Mouse