Lyophilization process optimization and molecular dynamics simulation of mRNA-LNPs for SARS-CoV-2 vaccine

Abstract

Some studies have shown that lyophilization significantly improves the stability of mRNA-LNPs and enables long-term storage at 2–8 °C. However, there is little research on the lyophilization process of mRNA-lipid nanoparticles (LNPs). Most previous studies have used empirical lyophilization with only a single lyoprotectant, resulting in low lyophilization efficiency, often requiring 40–100 h. In the present study, an efficient lyophilization method suitable for mRNA-LNPs was designed and optimized, shortening the total length of the lyophilization process to 8–18 h, which significantly reduced energy consumption and production costs. When the mixed lyoprotectant composed of sucrose, trehalose, and mannitol was added to mRNA-LNPs, the eutectic point and collapse temperature of the system were increased. The lyophilized product had a ginger root-shaped rigid structure with large porosity, which tolerated rapid temperature increases and efficiently removed water. In addition, the lyophilized mRNA-LNPs rapidly rehydrated and had good particle size distribution, encapsulation rate, and mRNA integrity. The lyophilized mRNA-LNPs were stable at 2–8 °C, and they did not reduce immunogenicity in vivo or in vitro. Molecular dynamics simulation was used to compare the phospholipid molecular layer with the lyoprotectant in aqueous and anhydrous environments to elucidate the mechanism of lyophilization to improve the stability of mRNA-LNPs. This efficient lyophilization platform significantly improves the accessibility of mRNA-LNPs.

The Effect of Nanoscale Modification of Nisin by Different Milk-Derived Proteins on Its Physicochemical Properties and Antibacterial Activity

“…Storage stability is an important indicator of the stability and reliability of a developed material during storage [ 62 ]. Generally, particle size and PDI are the key indicators to characterize the stability of nanoparticles [ 63 ], so the freshly prepared milk protein–nisin nanoparticles were stored at 25 °C and 4 °C for 28 days. The changes in particle size and PDI were monitored during the storage process, as well as the changes in visual appearance were recorded, and the results are shown in Figure 5 .…”

Single MVA-SARS-2-ST/N Vaccination Rapidly Protects K18-hACE2 Mice against a Lethal SARS-CoV-2 Challenge Infection

“…To further improve vaccine efficacy with regard to broader-protective and rapidly protective immunizations, several approaches have been undertaken. For mRNA vaccines, formulation and dosages have been optimized to result in a stronger activation of immune responses to improve the protective efficacy [ 49 , 50 , 51 , 52 , 53 , 54 ]. Several other approaches evaluated the effect of heterologous prime-boost regimens for the activation of broader-reactive and more robust immune responses [ 55 , 56 , 57 , 58 ].…”