Since the initial demonstration of gene expression in vivo through naked DNA injection, DNA vaccines, and gene therapies have advanced significantly. Plasmid DNA is used in both approaches to treat diseases by inducing protein expression. In comparison to virus-based vaccines, DNA vaccines offer high safety, low carrier-induced immunogenicity, and ease of production.

With more DNA vaccines advancing to clinical trials, the demand for high-purity pDNA escalates. However, scaling up DNA vaccine production faces challenges. Today, we delve into DNA vaccine production across different scales.

pDNA Production: Scale and Quality

The production of pDNA encompasses vector construction, cell bank preparation, fermentation, and purification. Quality assessment is based on supercoiled (SC) percentage, purity, stability, and potency. Large-scale production confronts the dual challenges of impurity removal and yield enhancement, with downstream processing posing a significant bottleneck. After cell harvest and lysis, techniques like Expanded-Bed Adsorption are used to initially capture pDNA. While chromatography dominates purification, its high cost and inefficiency have prompted the exploration of economical alternatives such as selective precipitation. Optimizing downstream processes to reduce costs, enhance purity, and increase yields is crucial for DNA vaccine production.

Small-Scale Production: Small-scale trials often utilize shake flasks, abandoning traditional methods in favor of efficient purification kits. Diverse lysis methods are employed, and purification often incorporates chromatography. The AEX/ethanol precipitation method is simple and cost-effective but limited to gram-scale applications. The SEC/TAC/AEX process, while capable of reaching gram-scale production, is costly and time-consuming.

Pilot-Scale Production: Yaohai Bio-Pharma has over a decade of experience in the microbial expression of pDNA. Yaohai has also established a GMP-level pilot-scale production platform, allowing it to meticulously balance yield, purity, and cost by leveraging state-of-the-art bioreactors and a diverse array of downstream technologies. The AEX–EBA/SEC approach achieves unparalleled purity, while also acknowledging its inherent complexity. In contrast, EBA offers enhanced processing capacity, albeit with a slight compromise in purity. Through this strategic blend of technologies and expertise, Yaohai Biotech stands as a reliable partner in advancing the frontier of pilot-scale production for CDMO projects.

Large-Scale Production: Large-scale purification confronts multiple challenges, with thermal lysis and membrane technologies enhancing efficiency but requiring further refinement for clinical-grade purity. Chromatography remains the cornerstone of downstream purification, often combined with non-chromatographic methods for optimal results. ATPS shows potential but requires validation at larger scales. Various purification pathways have their pros and cons, with AEX membranes/HIC/TFF emerging as highly efficient and versatile options, already proven in the production of multiple DNA vaccines.

Conclusion

DNA vaccine technology has been improved, resulting in enhanced production capacity and purity. However, current detection technologies and standardization remain issues. We anticipate ongoing technological developments that will address these challenges to further improve the manufacturing process of DNA vaccines. Yaohai can complete the amplification, optimization, and transfer of plasmid DNA production processes with high quality, delivering the drug substance and a comprehensive pilot-scale report to clients, thereby advancing the development process of CDMO projects. Yaohai is your trusted partner in this vast pDNA market.

Yaohai Bio-Pharma is also actively seeking institutional or individual global partners and offers the most competitive compensation in the industry. If you have any questions, please feel free to contact us: [email protected]

For more details about Yaohai Bio-Pharma, please kindly refer to the website: www.yaohaibio-pharma.com