中国生物工程杂志

The pcD-awte candidate malaria DNA vaccine could induce protective immune-responses through animal test, so it is necessary to establish a suitable preparative protocol with which the vaccine can be applied in clinical test as soon as possible. Here we reported an efficient preparative method, with which plasmid DNA vaccine could be produced in high yield and purity. Escherichia coli DH5α containing recombinant plasmid pcD-awte was grown through fed-batch fermentation in 5L fermentor with basic culture media of improved TB. After fermentation, the crude plasmid DNA　was obtained by alkaline lysis and the plasmid DNA was further purified with a three-step efficient chromatographic procedure, including Sepharose 6FF size exclusion chromatography , Plasmidselect thiophilic aromatic chromatography and Source 30Q anion exchange chromatography. The purified plasmid DNA was identified, which showed that the yield of final product reached 43.9 mg/L and that the quality met the standards of pharmaceutical-grade plasmid DNA.