mRNA Purification Focus Day - Tuesday, September 17
8:00 am Morning Check In & Coffee
8:50 am Chair’s Opening Remarks
Optimizing Upstream Synthesis to Simplify Purification Processes & Obtain High Quality mRNA
9:00 am Supercharging IVT Reactions for Co-Transcriptional Removal of dsRNA to Produce High Purity mRNA & Simplify Post-Translation Steps
Synopsis
- Optimizing IVT reactions for co-transcriptionally reducing dsRNA production to produce high purity mRNA
- Discussing strategies to rate-limit UTP feeding into the reaction to maintain yield while increasing purity
- Balancing multiple parameters like enzyme efficiency, reaction rate and raw material feeding for optimal IVT reactions
9:30 am Panel Discussion: Simplifying the Purification Process: Optimizing for High Quality DNA & mRNA Synthesis to Reduce Impurities
Synopsis
- Identifying critical parameters during DNA production that can enable simpler and more cost-efficient downstream purification of DNA
- How to optimize pre-purification processes, such as IVT reactions, to increase purity and organically reduce purification steps and time
- How do the above optimizations to DNA and mRNA synthesis change when considering circRNA?
10:30 am Morning Break & Networking
Enhancing dsRNA Purification & Characterization Through Chromatography for High Quality Drug Substance Production
11:30 am Improving Binding Capacity & Throughput of Therapeutic mRNA with Oligo dT (OdT) Immobilized Fibro Prototype Chromatography Media
Synopsis
- Evaluation of two commercially available OdT matrices and one prototype media for affinity chromatography of mRNA vaccine candidates
- Assessing OdT membrane for purification of mRNA vaccine candidates presented in recent manuscript
- Showcasing the impact of improved binding capacity and throughput afforded by novel OdT affinity chromatography media type
12:00 pm Panel Discussion: Boosting Assay Development for Purification Processes to Accurately Measure & Characterize dsRNA to Improve Drug Substance Safety
Synopsis
- Understanding dsRNA impurity profiles and assessing the best standards by which to characterize it
- Optimizing current cell-based assays to assess immunogenicity with greater sensitivity evoked by dsRNA
- Adopting and selecting appropriate process controls to enhance purification assays for dsRNA characterization
1:00 pm Lunch Break & Networking
Supercharging Purification from pDNA to mRNA Drug Product Without Compromising Yield to Achieve Desired Quality
2:00 pm Navigating DNA Linearization in its Supercoiled Form & Topology to Enhance DNA Purification Strategies
Synopsis
- Discussing how to optimize DNA linearization in its supercoiled form for faster and more cost effective pDNA isolation
- Assessing methods to differentiate between supercoiled and linear DNA with PLC assays
- Addressing the effect of initial plasmid purity on linearization efficiency and subsequent purifications
2:30 pm Highlighting Scale Up Considerations for Purification Processes to Maintain mRNA Purity & Integrity
Synopsis
- Maintaining parameters and controls when scaling up from small to large batches to ensure purification processes achieve desired mRNA yield and quality
- Ensuring novel resins meet standards for clinical grade material production
- Implementing scalable purification strategies into a GMP setting for cost-effective, efficient and GMP-compliant processes
3:00 pm Decontamination of Purified mRNA Vaccines & Therapeutics
Synopsis
Small amounts of contaminants remain in purified vaccines and are often the cause of a range of uncomfortable (but not deadly) immunogenic responses
We present a methodology that, in principle, will allow one to remove selected contaminants from purified mRNA vaccines and therapeutics with fast, efficient and scaleup bioprocessing
Our approach combines 2 steps:
- discovery and development of peptide affinity ligands that bind to a selected contaminant in preference to ss-RNA (desired product) or vice versa, and
- grafting and testing of these ligands to microporous synthetic polymer membranes to determine their binding affinity and selectivity in static and dynamic process modes
3:30 pm Rapid Screening of Materials of Contact for Scale-Up of mRNA-LNP Manufacturing
Synopsis
- Early screening of leachables and extractables during scale-up can reduce challenges with technical transfer and manufacturing
- As the process is scaled, the material contact time can increase several fold which would increase levels of E&L in your drug product
- We have developed a strategy for rapid screening of E&L to facilitate selecting materials of contact