Plasmid DNA fermentation strain and process-specific effects on vector yield, quality, and transgene expression.

摘要: Industrial plasmid DNA manufacturing processes are needed to meet the quality, economy, and scale requirements projected for future commercial products. We report development of a modified fermentation copy number induction profile that increases gene vaccination/therapy vector yields up 2,600 mg/L. determined that, in contrast recombinant protein production, secretion metabolic byproduct acetate into media had only minor negative effect on replication. also investigated impact differences epigenetic dcm methylase-directed cytosine methylation transgene expression, immunogenicity. While Escherichia coli production yield quality unaffected, dcm- versions CMV CMV-HTLV-I R promoter plasmids increased expression human cells. Surprisingly, despite improved is less immunogenic. Our results demonstrate it critical lock pattern (i.e., strain) early product strains may be superior therapy applications wherein reduced immunogenicity desirable vitro transient transfection such as AAV where beneficial.