Delivering nucleic acids using lipid nanoparticles (LNPs) to target cells has tremendous potential to enable new cell and gene therapies. In this presentation, we will discuss a platform approach of novel ionizable lipids and off-the-shelf reagents to encapsulate and deliver mRNA to the site of activation. In addition, we will share insights into microfluidic manufacturing technologies that accelerate scale-up by enabling downstream process development at the bench scale using a self-amplifying RNA (saRNA)-LNP vaccine as a case study. Furthermore, to exemplify the benefits of LNPs in the production of cell-based cancer therapies, we demonstrate a robust method for multi-step engineering of gene-edited CAR T cells.

Speaker:

Dr. Martin Rabel
Senior Field Application Scientist
Precision NanoSystems

The talk will present case studies of leveraging the power of at-line HPLC monitoring for process development to increase purity and yield of mRNA production. Approaches to optimize plasmid purification and linearization, IVT reaction and mRNA purifi cation will be described, including purification methods to achieve >95% supercoiled plasmid purity, optimize linearization reaction, and increase of IVT reaction yield from 3-5 mg/mL to 10-12 mg/mL. Further, use of at-line anayltics to monitor automated, continuous production in single use bioreactors to produce multi-gram amounts of mRNA will be demonstrated. Oligo dT-based affinity purification will be described with approaches to optimize dynamic binding capacity to reach up to 6 g/L. Multimodal chromatography and reverse-phase chromatography cases will be discussed for capture of non-polyadenylated RNA, and removal of dsRNA.

Speaker:

Rok Sekirnik
Head Process Development mRNA/pDNA
Sartorius BIA Separations, d.o.o.

This session will provide an overview of the extraordinary potential of mRNA technology for applications in the vaccine field, illustrate real-life examples of pre-clinical and clinical studies, and indicate new avenues for future developments.

Speaker:

Paolo Lusso, MD, PhD

National Institutes of Health  

The success of mRNA-based COVID-19 vaccines has triggered a rapid expansion of mRNA vaccines and therapeutics. An essential part of any mRNA therapeutic design is the 5’ cap structure, which is critical to the stability and expression of an mRNA. Learn how next-generation capping strategies impact manufacturing costs, timelines, and complexity. In addition, we will address considerations for process development, synthesis, and scale-up to streamline the mRNA manufacturing process.

Speaker:

Evan Myers
Associate Director, Process Development and Custom Development
TriLink BioTechnologies 

This session will provide an overview of commonly used physicochemical and biophysical methods used to characterize lipid nanoparticles (LNPs). After attending this session, you will have gained knowledge of commonly used techniques for LNP characterization, including an understanding of how the techniques work and what information that can be obtained. The session will provide examples on how these methods can inform us on the lipid distribution in LNPs, which we have used to guide how to vary their size and surface composition to increase protein production. The session will also cover some new and less commonly used techniques in the field currently being explored to characterize LNPs.

Speaker:

Gustav Emilsson, PhD.
Senior Scientist
AstraZeneca