Pilot-Scale Plasmid Vectors

The VDL offers pilot-scale plasmid produced in compliance with GLP. The pilot-scale production method yields 200 mg of purified, clean plasmid.

Briefly, overnight cultures are grown and inoculated into 20 liters of media in the fermenter. After 15-24 hours at 37ºC, the cells are removed by low speed centrifugation and treated with lysis solutions to break open the cells. After filtration through a 70 µm cell strainer, the lysate is aseptically transferred to a concentrator and the sample reduced to 1/10-1/15 of the original volume. Plasmid isolation is performed using Plasmid Select (Amersham Bioscience), a selective capture system in a three step process for purifying the supercoiled, covalently closed circle (ccc), form of plasmid DNA. This process involves: 1) RNA removal and buffer exchange by group separation on Sepharose™ 6 Fast Flow 2) Selective capture of supercoiled plasmid DNA with PlasmidSelect 3) Endotoxin removal and sample concentration on SOURCE™ 30Q. This purification is performed on an AKTA Explorer FPLC. Several quality control assays are performed on the DNA to ensure its identity, sterility and removal of endotoxins. Identity testing is by restriction enzyme analysis.