Slide 1: Bridging Small & Medium-Scale DNA Production
Bridging Small & Medium- Scale DNA Production: Practical Strategies for Scalable Manufacturing with High Quality DNA Process Development & Manufacturing Summit Boston, MA March 26, 2026 © 2023 IMUNON, Inc. PAGE 1
Slide 2: Topics
Topics • Plasmid Manufacturing scale-up case study: strategy and lessons learned from a well-controlled process resulting in high quality and competitive yield • Value of in-process testing within development plan to build a more robust, scalable process • Practical considerations to enable rapid de
Slide 3: Imunon's Manufacturing Story for High Quality Plasmid DNA-based Therapy
Imunon’s Manufacturing Story for High Quality Plasmid DNA-based Therapy • To support the company’s pDNA vaccine program, a small internal team produced >100 pDNA constructs at gigaprep scale • To enable multiple in vivo studies from single pDNA batch, fed-batch fermentation and small-scale, highly i
Slide 4: Lead Candidate – phIL-12 Non-viral Gene Therapy
Lead Candidate – phIL-12 Non-viral Gene Therapy Status: Enrolling Phase 3 (OVATION 3; randomized, 500-patient trial) © 2023 IMUNON, Inc. PAGE 4
Slide 5: Motivation for Internal Development of Manufacturing Capabilities
Motivation for Internal Development of Manufacturing Capabilities • After several years of reliance on external CDMO partners for clinical API supply, where prices were consistently increasing, we projected that the COGS would prove to be unaffordable if we did not take action • Manufacturing capabi
Slide 6: pDNA Manufacturing Process Requirements
pDNA Manufacturing Process Requirements • In evaluating potential of internal manufacturing capabilities to meet the challenge, the process must be: Efficient, balancing cost and productivity Easily scaled to sufficient batch sizes, maintaining high quality and control Ideally fully enclosed,
Slide 7: pDNA Manufacturing Process Summary
pDNA Manufacturing Process Summary Key Aspect Description Nucleic Acid pDNA End Use API Biomass Production Fed-Batch Microbial Fermentation E. coli Stbl2 (Small Scale-vaccine) Strain E. coli DH10B (Medium Scale-gene therapy) DSP Method Alkaline Lysis © 2023 IMUNON, Inc. PAGE 7
Slide 8: Primary Considerations
Primary Considerations At the outset, what were the key steps and associated questions? 1) How can the lysis system be scaled? • What single-use assembly configurations are available? • If none are available, could similar assemblies be modified to specifications? 2) What depth filter train surface
Slide 9: USP/Fermentation Scale-Up
USP/Fermentation Scale-Up 10L to 100L • Staying with same fermenter manufacturer reduces reactor design & control variability • Remaining development work is simpler – Determine actual max fermentation volume – Feed pump calibration & feed rate scaling – Sterile feed medium production, including SUA
Slide 10: General Approaches to Scaling Lysis
General Approaches to Scaling Lysis • Design process that is modular or utilizes components that are readily available in different sizes – Similar to chromatography, fix one or two dimensions, vary third to meet requirements • Measure as many system parameters as possible • Calibrate equipment prio
Slide 11: General Approaches to Scaling Clarification
General Approaches to Scaling Clarification • Disc-type centrifuges can be cost-effective if available • Depth filter trains are effective but can be the most expensive DSP component per batch – Vendors often offer on-site development support to aid in filter selection – Number of filters in stack a
Slide 12: General Approaches to Scaling UFDF
General Approaches to Scaling UFDF • General principles of TFF processing of pDNA steams should be followed − Membrane surface area chosen to enable reasonable processing time − Crossflow rate must be high enough to prevent membrane fouling − Membrane flux should be optimized • Optimal membrane flux
Slide 13: General Approaches to Scaling Chromatography
General Approaches to Scaling Chromatography • Generally, most straight-forward DSP unit operation to scale up • When scaling, keep column height constant and increase column diameter • Adjust flow rate to stay within pressure limits of packed column • Approach is typically bind-and-elute – Mobile p
Slide 14: In-Process Testing/PAT Strategy to Build Robust, Scalable Process
In-Process Testing/PAT Strategy to Build Robust, Scalable Process • Early on, sample and retain everything (buffers, process steps, pre-/post-filtration, waste, etc.) • Establish in-process acceptance criteria when data support them (flexibility is critical early on) • In continuous processes, use s
Slide 15: In-Process Testing/PAT Strategy to Build Robust, Scalable Process
In-Process Testing/PAT Strategy to Build Robust, Scalable Process • Consider fraction volumes and reality of dilution when setting in-process specs/strategy – For example, gDNA is a troublesome impurity in pDNA process streams – We may be tempted to: • Set in-process spec the same as final product (
Slide 16: In-Process Testing/PAT Strategy to Build Robust, Scalable Process
In-Process Testing/PAT Strategy to Build Robust, Scalable Process • However, being too conservative may cause unnecessary pDNA loss • See the single batch below, purified by AEC over two cycles (identical loads) – In Cycle 1, early fraction volume is low (0.3 L) to select out fractions high in gDNA
Slide 17: Practical Considerations to Enable Rapid Development of a Well-Controlled Process
Practical Considerations to Enable Rapid Development of a Well- Controlled Process with High Quality Within Budget • Project annual material/supply needs, communicate to vendors – Develop key relationships into partnerships • Determine if early-DSP steps require highest purity reagents, sterile SUAs
Slide 18: Thank you!
Thank you! © 2023 IMUNON, Inc. PAGE 18