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- Title
318. Collaborative Development of an Effective Method To Produce Clinical RNAi Products.
- Authors
Ramakrishnan, Sriram; Darnell, Jeffrey E.; Gates, Gary; Gonzales, Hugo L.; Miller, Justin; Hebel, Henry
- Abstract
The demand for high purity plasmid DNA for use in the fields of gene therapy and DNA vaccination is escalating rapidly. The new and expanding field of RNAi also relies on plasmids to express short double-stranded RNA molecules that specifically target mRNAs and degrade them in a sequence-dependent manner for therapeutic purposes. Plasmid-based approaches allow the use of multiple siRNAs that target multiple genes of a disease pathway or of viral pathogens. While this represents an advantage for the RNAi technology, it adds challenges to the development of a production process. Only through close collaboration were the teams at ADViSYS, Inc. and Nucleonics, Inc. able to develop a method suitable for clinical production.Based on the previous history and technology available to both groups, a number of hybrid methods were discussed and attempted. After several rounds of meetings and experiments, a final method was selected. The method takes full advantage of ADViSYS's scaleable lysis and membrane chromatography and adds a polishing column specified by Nucleonics to enrich supercoiled plasmid and provide additional contaminant removal. Through the use of this method, three ADViSYS personnel are able to produce for Nucleonics over 10gm of final pure bulk plasmid DNA from a single 100L fermenter within 1 week. The quality of material exceeded the client's expectations, with greater than 99% purity for the two large scale batches produced to date. This method was also able to consistently enrich the total supercoiled content by 11 to 14%.We believe this development work further validates ADViSYS's core processing technologies and the concept that initial flexible process steps are required to produce high quality plasmid-based products for a wide variety of indications and routes of delivery. The collaborative nature of this program also represents a strategic advantage for both organizations as RNAi products will continue to expand and several will enter clinical trials.Molecular Therapy (2006) 13, S121–S121; doi: 10.1016/j.ymthe.2006.08.374
- Subjects
DNA; GENE therapy; VACCINATION; SMALL interfering RNA; PATHOGENIC microorganisms; NUCLEONICS Inc.; ADVISYS Inc.; CLINICAL trials
- Publication
Molecular Therapy, 2006, Vol 13, pS121
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1016/j.ymthe.2006.08.374