Conference Day One | Wednesday, March 26, 2025
7:50 am Check In & Light Breakfast
8:50 am Chair’s Opening Remarks
Enhancing DNA Fidelity & Quality by Measuring Impurities to Increase Yield of High-Value DNA for Potent Therapies
9:00 am Impacts of Plasmid Product Quality Attributes on mRNA PolyA Tail Length
Synopsis
- Understand the importance of pDNA as a starting material on mRNA product quality to inform plasmid design
- Navigating DNA polyA tail length heterogeneity to guide upstream process development decisions
- Engineering a plasmid that impacts polyA tail length to maximize product quality
9:30 am Optimizing Plasmid DNA Clarification to Maximize DNA Purity
Synopsis
- Evaluating different parameters to minimize key DNA impurities: host cell DNA and open circular DNA
- Exploring alternative clarification unit operations to enhance impurity removal and increasing yield
- Understanding key scaling parameters during lysis and clarification operations
10:00 am Cell-Free DNA Technology: Next Generation Solutions for mRNA Manufacturing
Synopsis
- A cell-free, enzymatic DNA manufacturing process to accelerate linDNA template production
- Reducing linDNA template production timelines by half, enabling faster mRNA synthesis
- Eliminating bacterial cells from the amplification process
10:30 am Morning Break & Speed Networking
Synopsis
Our dedicated speed networking session is the perfect opportunity to have in-depth conversations and forge long-lasting connections with fellow technical experts working with plasmid and synthetic DNA
Refining Upstream Process Development & Fermentation to Maximize Downstream Recovery & DNA Stability for Higher Efficiency Advanced Therapeutics
11:30 am Optimizing Fermentation Conditions for Robust Plasmid Production
Synopsis
- Reducing fermentation stresses for robustness of plasmid production
- Balancing growth and yield for plasmid production to maximize efficiency
- Evaluating media conditions for optimal growth and plasmid recovery
12:00 pm Pure Synthetic DNA: Unlocking the Genetic Medicines Needs
Synopsis
- Quality comes first – Ensuring high quality products and reproducible results
- Faster Delivery Time – Accelerating access to DNA material through high performance manufacturing process
- Scalability – Meeting any amount demanded: from R&D and clinical to commercial
12:15 pm Debottlenecking Vaccine Manufacturing; Simplifying DSP Process
Synopsis
- Optimizing host strain, plasmid design, media composition and fermentation conditions for high biomass, plasmid yield and quality, facilitating downstream purification of ScpDNA
- pDNA recovery is a critical step in the downstream process, removing most contaminants while concentrating pDNA
- Pharmaceutical pDNA has stringent requirements of purity, homogeneity, and efficacy, and SC pDNA is the most stable, efficient, and biologically active
12:45 pm Lunch & Networking
Elevating Effective Lysis & Purification of DNA to Increase Downstream Efficiency & Yield to Maximize Manufacturing Efficiency
1:45 pm Thinking Beyond Traditional Resin Based Chromatography for Improved Purification & DNA Quality
Synopsis
- Exploring the possibilities of monolithic resins, membrane absorbers and ion exchange chromatography to effectively purify pDNA
- How to maintain cost-effectiveness whilst employing advanced purification methods to ensure sustainable DNA production
2:15 pm Optimization of Plasmid DNA (pDNA) Extraction via Lysis to Enhance Manufacturing Efficiency and QualitY
Synopsis
Plasmid DNA (pDNA) isolation involves a crucial cell lysis step using a strong alkaline solution, followed by neutralization and precipitation to recover target pDNA while removing cellular contaminants like genomic DNA and proteins. Uncontrolled alkaline use and mechanical stress can degrade pDNA, and highly viscous solutions may cause heterogeneities or require extensive mixing, further compromising pDNA integrity.
Taking these aspects into consideration, we will present challenges and optimization opportunities in the context of scaling-up pDNA process for mRNA applications. Key considerations include maximizing product yield and quality while minimizing residual impurities, managing product/waste volumes and ensuring process step robustness.
As part of our evaluation of pDNA extraction methods, we will present an automated in-line lysis system (Alkalizator from Sartorius BIA Separations) with a closed single-use loop, tailored for process development up to pilot-scale within a GMP environment. It features advanced mixing control, enabling automatic, continuous processing of up to 500 liters of resuspended cells per workday, depending on configuration.
We will demonstrate the system's application in pDNA process, showcasing in-line lysis results including impact on pDNA quality.
Key Learning Objectives:
- Comprehend the challenges associated with alkaline lysis in pDNA extraction
- Evaluate the advantages of the Sartorius in-line lysis system in comparison to traditional batch lysis methods
- Explore at-line in-process control strategies tailored for managing complex crude lysis samples.
This work was funded by Sanofi. The authors may hold shares and/or stock options in their respective companies.
2:45 pm Panel Discussion: Optimizing Downstream DNA Process Development through Novel Lysis & Purification Methods to Protect Yield & Quality
Synopsis
- How to streamline downstream process development and employ the latest methods and technologies to maximize efficiency of DNA production
- How to reduce the manufacturing timeline and maintain cost efficiency whilst producing high quality starting materials and API
- Understanding the regulatory landscape of lysis and purification of DNA for use in advanced therapies
3:30 pm Afternoon Break & Poster Session
Synopsis
This is your opportunity to share your innovations and breakthroughs with your DNA manufacturing peers
Integrating Analytical Testing & Quality Control Methods to Streamline Release of Manufactured DNA for High Quality mRNA, Cell & Gene Therapies & Vaccines
4:30 pm Analytical Considerations for Plasmid DNAs as Starting Material for Cell & Gene Therapy Products in Clinical Development
Synopsis
- Outlining the characterization and quality control of plasmid DNAs
- Employing HPLC, UV and NGS tools to test quality attributes to understand pDNA purity
- Stability behaviour and shelf-life setting for long-term storage to maximize use of pDNA and advanced therapies
5:00 pm Accelerating & De-risking the Path to Clinical Drug Development with the Cell-Free Synthesis of DNA
Synopsis
- Unlock greater speed and sequence diversity for screening by eliminating the time spent cloning plasmid DNA
- More economically scale DNA synthesis for personalized therapy
- De-risk clinical scale-up earlier in the development process
5:30 pm Ministring DNA: a Novel Linear DNA Minivector with Superior Fidelity & Enhanced Stability for Gene Therapy Applications
Synopsis
- Enhancing stability of gene therapies with Ministring DNA through resisting destabilization, maintaining structural integrity at critical pH levels, and ensuring consistent non-viral gene delivery
- How Trehalose-enabled lyophilization allows for room-temperature storage while maintaining functionality, facilitating use in low-resource settings
- Ensuring high fidelity, scalability and regulatory compliance through in vivo manufacturing in E.coli