Plasmid DNA Vaccines Manufacturing
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.
Related Technical Articles
- An overview of the cell harvest, lysis, neutralization, and clarification steps used for pDNA purification along with case study and practical advice.
- Get a step-by-step overview of the plasmid DNA manufacturing process and the challenges in pDNA downstream purification. Read our free pDNA downstream purification eBook for mRNA, plasmid-based DNA Vaccines, and Viral Vector applications.
- Cost Modeling Vaccine Manufacturing: Estimate Production Costs for mRNA and other Vaccine ModalitiesA custom-designed cost model is used to explore the economics of vaccine manufacturing across several different modalities including mRNA. The model enables greater process understanding, simulates bottlenecks, and helps to optimize production efficiency.
- Influenza vaccines are commonly made using egg-based and cell-based manufacturing strategies. Find step-by-step information on the manufacturing process for each method.
- mRNA technology uses non-viral delivery systems and offers a great deal of versatility. Delivery of an mRNA into the cytosol of a cell can induce the production of a target protein which can function as a therapeutic or prophylactic, act as an antigen to trigger an immune response for vaccination purposes, replace a defective protein or activate an anti-tumor response.
- Understand how mRNA vaccines induce immunity. and read how synthetic mRNA is prepared for vaccine immunogens and other biopharmaceuticals. Find reagents for synthesis of mRNA.
- This technical article breaks down the steps of upstream and downstream bioprocessing and formulation of virus-like particle vaccines.
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Find More Articles and Protocols
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.
Related Videos
Clarification
Achieve efficient, robust, clarification to maximize success of your downstream purification operations by reducing the levels of impurities and particulates.
Downstream - Tangential Flow Filtration
Achieve yield, efficiency and pDNA recovery goals while ensuring robust impurity removal
Bioprocessing Liquid Cell Culture Media & Buffers
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.
Downstream Chromatography
Tackle and overcome any pDNA chromatographic purification challenge in your downstream process to achieve greater productivity, efficiency and speed.
- Buffers & pH Ajusters
- Membrane Chromatography with Natrix® Q Pilot Chromatography Membranes
- Membrane Chromatography with Natrix® Q Recon Mini
- Capture an/or Polishing Chromatography with Fractogel® EMD DEAE Chromatography Resins
- Capture an/or Polishing Chromatography with Fractogel® EMD DMAE Chromatography Resins
Formulation, Sterile Filtration & Fill-Finish
Ensure patient safety with a reliable and robust sterile filtration process
Analytics Software & PAT Technology
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.
Product Services
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.
Related Resources
- Application Note: Cell Harvest, Lysis, Neutralization & Clarification of Plasmid DNA
- Application Note: Tangential Flow Filtration (UF/DF) of Plasmid DNA
How are ultrafiltration and diafiltration used in plasmid DNA manufacturing and how should you optimize this process? Learn more in our technical article and download our free eBook.
- Application Note: Chromatographic Purification of Plasmid DNA
See case study examples of how to optimize chromatographic purification of plasmid DNA for Biopharmaceutical Applications.
- Application Note: Sterilizing Grade Filtration Unit Operations for Plasmid DNA Processes
We discuss the final filtration step of plasmid DNA manufacturing and its key considerations and challenges. Download our free pDNA downstream purification eBook or ask our experts.
- White Paper: Designing a Plasmid DNA Downstream Purification Process, for mRNA
- Webinar: Scalable Purification of Plasmid DNA for use in Vaccine Manufacturing
- Webinar: Effective and Efficient Design of a Downstream Purification Process for Plasmid DNA
- Application Note: Scalable Purification of Plasmid DNA – Strategies & Considerations for Vaccine and Gene Therapy Manufacturing
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