IBMM, University of Montpellier, CNRS, ENSCM, Montpellier France.
Sanofi Pasteur, 1541 avenue Marcel Mérieux, 69280 Marcy l'Etoile, France.
Anal Chem. 2022 Mar 22;94(11):4677-4685. doi: 10.1021/acs.analchem.1c04778. Epub 2022 Mar 7.
Messenger RNA vaccines have come into the spotlight as a promising and adaptive alternative to conventional vaccine approaches. The efficacy of mRNA vaccines relies on the ability of mRNA to reach the cytoplasm of cells, where it can be translated into proteins of interest, allowing it to trigger the immune response. However, unprotected mRNA is unstable and susceptible to degradation by exo- and endonucleases, and its negative charges are electrostatically repulsed by the anionic cell membranes. Therefore, mRNA needs a delivery system that protects the nucleic acid from degradation and allows it to enter into the cells. Lipid nanoparticles (LNPs) represent a nonviral leading vector for mRNA delivery. Physicochemical parameters of LNPs, including their size and their charge, directly impact their behavior and, therefore, their cellular internalization. In this work, Taylor dispersion analysis (TDA) was used as a new methodology for the characterization of the size and polydispersity of LNPs, and capillary electrophoresis (CE) was used for the determination of LNP global charge. The results obtained were compared with those obtained by dynamic light scattering (DLS) and laser Doppler electrophoresis (LDE).
信使 RNA 疫苗作为一种有前途和适应性的替代传统疫苗方法，已经成为焦点。mRNA 疫苗的功效依赖于 mRNA 进入细胞细胞质的能力，在那里它可以被翻译成感兴趣的蛋白质，从而引发免疫反应。然而，未受保护的 mRNA 不稳定，容易被外切酶和内切酶降解，其负电荷被带负电荷的细胞膜静电排斥。因此，mRNA 需要一种输送系统来保护核酸免受降解并允许其进入细胞。脂质纳米颗粒 (LNP) 是 mRNA 输送的一种非病毒主要载体。LNP 的理化参数，包括其大小和电荷，直接影响它们的行为，因此也影响它们的细胞内化。在这项工作中，泰勒分散分析 (TDA) 被用作一种新的方法来表征 LNP 的大小和多分散性，而毛细管电泳 (CE) 则用于测定 LNP 的总电荷。所得结果与动态光散射 (DLS) 和激光多普勒电泳 (LDE) 得到的结果进行了比较。
