Biophysical characterisation of the structure of a SARS-CoV-2 self-amplifying - RNA (saRNA) vaccine

Abstract

The current SARS-Covid-2 pandemic has led to an acceleration of messenger ribonucleic acid (mRNA) vaccine technology. The development of production processes for these large mRNA molecules, especially self-amplifying mRNA (saRNA) has required concomitant development of analytical characterisation techniques. Characterising the purity, shape and structure of these biomolecules is key to their successful performance as drug products. This paper describes the biophysical characterisation of the Imperial College London Self amplifying viral RNA vaccine (IMP 1) developed for SARS CoV 2. A variety of analytical techniques have been used to characterise the IMP 1 RNA molecule. In this paper we use UV spectroscopy, dynamic light scattering (DLS), size exclusion chromatography small angle scattering (SEC-SAXS) and circular dichroism (CD) to determine key biophysical attributes of IMP-1. Each technique provides important information about the concentration, size, shape, structure and purity of the molecule.

Mesoscopic Structure of Lipid Nanoparticle Formulations for mRNA Drug Delivery: Comirnaty and Drug-Free Dispersions

“…Small-angle X-ray and neutron scattering (SAXS, SANS), dynamic light scattering (DLS, also named photon correlation spectroscopy, PCS), cryo- and freeze-fracture transmission electron microscopy (TEM), and atomic force microscopy (AFM) are, among other methods, widely used for the study of the mesoscopic structure of mRNA-LNPS. − Nevertheless, the internal structure of the mRNA-LNPS could by far not yet be resolved as good as the various published quite detailed but essentially artistic representations might suggest. ,, …”