To ensure effective in vivo delivery, mRNA vaccines rely on not only optimized antigen sequences but also a robust delivery system capable of overcoming extracellular barriers, facilitating endosomal escape, and evading intracellular immune responses. Lipid nanoparticles (LNPs) are currently the most widely utilized platform for this purpose. Areterna offers mRNA-LNP encapsulation services at a scale of milligrams, employing three established lipid formulations (SM102, ALC0315, MC3) as well as a diverse array of novel lipid compositions tailored to specific delivery requirements.
Schematic Diagram of Typical LNP Formula
LNPs are typically composed of four lipids – cationic or ionizable lipids, cholesterol, phospholipids, and PEG-lipids.
Image Source: Acta Biomater. 2021 Sep 1;131:16-40. doi: 10.1016/j.actbio.2021.06.023
Presentation of Data: Encapsulation of Fluc-eGFP mRNA with N1-Me-pUTP (5’CAP) Using SM102
1. mRNA-LNP Particle Size and PDI
Particle size and polydispersity index (PDI) play crucial roles in LNP biodistribution, cellular uptake, and, ultimately, therapeutic efficacy. Dynamic light scattering (DLS) was utilized to assess the particle size and distribution of the encapsulated mRNA depicted in Figure 1. After encapsulating Fluc-eGFP mRNA with N1-Me-pUTP (5’CAP) using SM102, the particle size measured 115.7 nm, with a PDI of 0.09, indicating a uniform and monodisperse population.
2. mRNA-LNP Encapsulation Efficiency
The encapsulation efficiency represents the percentage of mRNA effectively enclosed within the LNP, serving as a critical measure for assessing active ingredient loading. In Figure 2, the encapsulation rate of Fluc-eGFP mRNA with N1-Me-pUTP (5’CAP) was quantified using RiboGreen fluorescent dye on a spectrometer, surpassing 95%.
Summary
Areterna distinguishes itself as a comprehensive hub for mRNA research and development. Going beyond its role as a cost-effective supplier of raw materials, we empower researchers with a range of integrated solutions.
Our services go beyond standard mRNA preparation (at the mg-scale, achieving yields of 10 mg/ml). We offer customized sequence design and optimization, meticulously tailoring each component (including cap structures, 5’UTR, CDS, 3’UTR, Poly A, and nucleotides) to maximize efficacy. To facilitate effective delivery, we provide expert mRNA-LNP encapsulation services, offering a choice between established high-quality formulas (SM102, ALC0315, MC3) or the exploration of our diverse library of innovative lipid combinations.