Plasmid DNA Vaccines Manufacturing

Graphic of the plasmid DNA vaccine process

Plasmid DNA Vaccines Process

Plasmid DNA (pDNA) is an important component of viral vector therapies. These circular DNA molecules can be used as the therapeutic transgene, to code for the viral capsid or as the vaccine itself. DNA vaccines have been approved for use in animals and have been developed against the SARS-CoV-2 virus. pDNA is also used as the starting material for mRNA vaccines.

pDNA manufacturing presents several challenges. Production suffers from low productivity of microbial fermentation. Additionally, the purification process is complicated by the fact that the bacterial lysate is highly viscous and contains contaminants with properties similar to pDNA, leading to low resolution separation. These challenges can be overcome with use of an end-to-end platform consisting of advanced single-use technologies.

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Accelerate Time to Clinical while Increasing Upstream Productivity

pDNA presents several challenges that can be overcome with advanced single-use technologies for cell harvest and clarification. Centrifugation or microfiltration TFF (MF-TFF) is used to remove spent fermentation broth while depth filters have demonstrated excellent performance over a wide range of conditions for the clarification.

Achieve Yield and Efficiency Goals with Robust Impurity Removal

Anion exchange (AEX) has demonstrated robust clearance of proteins, RNA, gDNA and endotoxin. Plasmid isoforms, on the other hand, are very challenging to separate with ion exchange. In this case, HIC can be used and is well-placed following AEX due to the high salt eluate pool.

Maximize Downstream Recovery

Purification of pDNA is complex, requiring a combination of TFF and chromatography. TFF can be placed between the clarification and chromatography steps, where residual impurities can be washed through diafiltration and pDNA concentrated for chromatography. Both resins and membranes can be used for chromatographic purification. Resins offer flexible installations and good selectivity while membranes offer high binding capacity and flow rates.

Ensure Patient Safety

Once purified and formulated, pDNA must be sterile filtered to ensure patient safety. While this step may appear to be a relatively simple operation, filtration of pDNA can be challenging due to the large size of plasmids, high viscosity of the solution and bacterial retention for adjuvanted formulations. Many process parameters must be considered to optimize sterilizing grade filtration including salt concentration, plasmid size, purity, and plasmid concentration.

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We offer the industry’s highest quality sterile filtered liquid capabilities, supplying ready-to-use cell culture media, buffers, CIP and SIP products from GMP facilities worldwide to optimize your biopharma production.

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Process analytical technology (PAT) and analytics software can be used to build quality into the protein subunit vaccine manufacturing processes by monitoring and controlling processes inline and in real time. 

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We offer a variety of product services to meet the demands that your work requires, from systems and equipment installation, systems qualification, validation services, maintenance and service plans, training and specialized Services. 

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