Cimmino Wanda, Angelillo Alessia, Rea Giuseppina, Kalligosfyri Panagiota M, Nele Valeria, Campani Virginia, De Rosa Giuseppe, Cinti Stefano
Department of Pharmacy, University of Naples Federico II, 80131 Naples, Italy.
Department of Life Science, Health and Health Professions, Link Campus University, Rome 00165, Italy.
Anal Chem. 2025 Jun 17;97(23):11968-11973. doi: 10.1021/acs.analchem.5c00692. Epub 2025 May 24.
Lipid nanoparticles (LNPs) represent a versatile delivery platform proposed for a wide range of therapies based on nucleic acids, including microRNA (miRNAs). The ability of LNPs to encapsulate and protect RNA from degradation, as well as their ability to promote cellular uptake, has led to their clinical use with the approval of RNA-based medicinal products, i.e., COVID vaccines. In this context, a growing number of LNP formulations with improved transfection and biocompatibility are under development, requiring rapid, sensitive, and robust quality control tests, e.g., for the quantification of the encapsulated RNA. Nowadays, classical analytical approaches such as fluorescence, ultraviolet-visible (UV-vis) spectrophotometry, and chromatography are mainly used for the quantification of the encapsulated drug. However, the user-friendly and cost-effective quantification of the encapsulation efficacy within LNPs represents an important research focus, as it would allow monitoring of the amount of encapsulated RNA, thus providing immediate quality control. In this work, we present the adaptation of an electrochemical strip to quantify the encapsulation of a miRNA, i.e., miR-218, whose antitumor effect has been widely reported in the literature within LNPs. We provide a rapid and sensitive method to assess the concentrations of miRNA actually encapsulated, obtaining satisfactory agreement compared to the traditional fluorimetric approach. Specifically, the platform is based on a commercial gold-screen-printed electrode modified with a DNA probe designed to be fully complementary to the target miRNA-218. The electrochemical system was successfully combined with a 3D-printed chamber that allowed the use of multiple electrodes simultaneously and the use of Triton X-100 surfactant to disrupt the LNPs and release the encapsulated miRNA-218 achieving a detection limit as low as 1 nM.
脂质纳米颗粒（LNPs）是一种多功能递送平台，被提议用于基于核酸的多种疗法，包括微小RNA（miRNAs）。LNPs能够封装并保护RNA不被降解，以及促进细胞摄取的能力，使得它们在基于RNA的医药产品（即新冠疫苗）获批后得以临床应用。在此背景下，越来越多具有改进转染和生物相容性的LNP制剂正在研发中，这需要快速、灵敏且稳健的质量控制测试，例如用于定量封装的RNA。如今，诸如荧光、紫外可见（UV-vis）分光光度法和色谱法等经典分析方法主要用于定量封装的药物。然而，对LNPs内封装效率进行用户友好且经济高效的定量是一个重要的研究重点，因为这将允许监测封装的RNA量，从而提供即时质量控制。在这项工作中，我们展示了一种电化学试纸条的适配，用于定量一种miRNA（即miR-218）的封装，其抗肿瘤作用在文献中已被广泛报道于LNPs内。我们提供了一种快速且灵敏的方法来评估实际封装的miRNA浓度，与传统荧光法相比获得了令人满意的一致性。具体而言，该平台基于一个用DNA探针修饰过的商业金丝网印刷电极，该DNA探针设计为与目标miRNA-218完全互补。电化学系统成功地与一个3D打印腔室相结合，该腔室允许同时使用多个电极，并使用Triton X-100表面活性剂破坏LNPs并释放封装的miRNA-218，实现了低至1 nM的检测限。
