Development of Purification Process of Plasmid DNA for Gene Therapy by Introducing Affinity Membrane Filtration

2015 Fiscal Year Final Research Report

• Project/Area Number: 25420800
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Properties in chemical engineering process/Transfer operation/Unit operation
• Research Institution: Nagoya University
• Principal Investigator: KATAGIRI Nobuyuki 名古屋大学, 工学(系)研究科(研究院), 助教 (00345919)
• Co-Investigator(Kenkyū-buntansha): IRITANI Eiji 名古屋大学, 大学院工学研究科, 教授 (60144119)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: プラスミドDNA / 精製 / アフィニティ / 膜濾過 / リガンド / 吸着 / 脱着 / 菌体破砕

Outline of Final Research Achievements

The development of a purification process of medical products purity plasmid DNA by affinity membrane filtration was examined. From the results of particle size distribution, photomicrograph, membrane filtration of slurry, and electrophoretic analysis of nucleic acids, it was found that the introduction of electroporation was useful for elution of plasmid DNA from bacterial cells. The iron oxide particle was selected as a ligand for two-stage affinity microfiltration of plasmid DNA. In the first stage microfiltration, the experiment was conducted under the condition of pH 5, and the cake of iron oxide particle with plasmid DNA bound was obtained and the impurities were removed. Next, water (pH 9 10 and 12) permeations through the cake were performed to desorb the bound plasmid DNA. It is revealed that the degree of purification of nucleic acid is high from the analysis of the recovery solution, the usefulness of this method has been confirmed.

Free Research Field

工学・プロセス工学