Optimization of self-amplifying RNA and mRNA production and formulation through advanced microfluidics

This project aims to systematically enhance the structure, synthesis, and formulation of sa-RNA and mRNA (sa-mRNA).

First, we will investigate NSPs of other alphaviruses to improve the efficiency of the non-structural proteins (NSPs) derived from the Venezualan Equine Encephalitis virus (VEEV) in our current sa-RNA and examine their intrinsic immunogenicity and its effect on vaccination potential. Second, by SBO project with a primary economic finality aimed at the transfer to existing companies modifying the primary structure, we aim to enhance sa-mRNA stability, which could impact both in vivo immunogenicity and the dose needed for vaccination.

While mRNA vaccine production has primarily focused on scaling up, there is a growing need for small-scale, rapid, and flexible production capacities due to the rise of personalized medicine. A significant bottleneck in producing mRNA-based vaccines is the requirement for large volumes and the time needed to switch between different sequences. We plan to address these issues using advanced microfluidic techniques to optimize both enzymatic RNA synthesis, purification and the formulation of RNA in nanoparticles for in vivo delivery.

Using a funnel-based selection procedure, we will evaluate the most promising vaccine candidates in vitro and in relevant in vivo models to evaluate their chemical properties, intracellular degradation kinetics, uptake capacity, expression levels, biodistribution and immunogenicity. We will use wellestablished mouse tumor models such as B16 and CT26 with defined neo-epitopes as a model for a personalized medicine in oncology. Given the importance of rapidly adapting vaccine formulations in response to emerging variants of concern in infectious diseases, we will in future also explore candidate vaccine formulations against new infectious diseases.