使用微流控装置生产载 siRNA 的脂质纳米粒。
Production of siRNA-Loaded Lipid Nanoparticles using a Microfluidic Device.
机构信息
Division of Applied Chemistry, Faculty of Engineering, Hokkaido University; JST PRESTO;
Graduate School of Chemical Sciences and Engineering, Hokkaido University.
出版信息
J Vis Exp. 2022 Mar 22(181). doi: 10.3791/62999.
The development of functional lipid nanoparticles (LNPs) is one of the major challenges in the field of drug delivery systems (DDS). Recently, LNP-based RNA delivery systems, namely, RNA-loaded LNPs have attracted attention for RNA therapy. In particular, mRNA-loaded LNP vaccines were approved to prevent COVID-19, thereby leading to the paradigm shift toward the development of next-generation nanomedicines. For the LNP-based nanomedicines, the LNP size is a significant factor in controlling the LNP biodistribution and LNP performance. Therefore, a precise LNP size control technique is indispensable for the LNP production process. Here, we report a protocol for size controlled LNP production using a microfluidic device, named iLiNP. siRNA loaded LNPs are also produced using the iLiNP device and evaluated by in vitro experiment. Representative results are shown for the LNP size, including siRNA-loaded LNPs, Z-potential, siRNA encapsulation efficiency, cytotoxicity, and target gene silencing activity.
功能性脂质纳米粒（LNPs）的开发是药物传递系统（DDS）领域的主要挑战之一。最近，基于 LNPs 的 RNA 传递系统，即负载 RNA 的 LNPs，已引起人们对 RNA 治疗的关注。特别是，负载 mRNA 的 LNP 疫苗已被批准用于预防 COVID-19，从而导致向开发下一代纳米药物的范式转变。对于基于 LNP 的纳米药物，LNP 的大小是控制 LNP 生物分布和 LNP 性能的重要因素。因此，精确的 LNP 尺寸控制技术对于 LNP 的生产过程是必不可少的。在这里，我们报告了一种使用微流控装置（称为 iLiNP）进行尺寸控制的 LNP 生产的方案。还使用 iLiNP 设备生产了负载 siRNA 的 LNPs，并通过体外实验进行了评估。展示了 LNP 尺寸的代表性结果，包括负载 siRNA 的 LNPs、Zeta 电位、siRNA 包封效率、细胞毒性和靶基因沉默活性。
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