Drennan Brady W, Schug Kevin A
Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX, USA.
Anal Bioanal Chem. 2025 Sep 13. doi: 10.1007/s00216-025-06096-4.
Gene therapies are rapidly advancing as drug modalities, serving as a means to treat previously undruggable pathways. The use of small interfering ribonucleic acid (siRNA), messenger RNA (mRNA), and guide RNA (gRNA) in clustered regularly interspaced short palindromic repeats (CRISPR)-based gene editing, combined with lipid nanoparticles (LNPs), has demonstrated effective drug delivery. These complex delivery systems often require multiple analytical methodologies to achieve comprehensive characterization, some of which remain underdeveloped or inadequately adapted for RNA-LNP formulations. Commonly used batch-based methods, such as dynamic light scattering (DLS) or the modified RiboGreen assay, are frequently hindered by sample heterogeneity, a limitation that can be addressed through analytical separations. This review discusses the challenges limiting analytical separations for RNA-LNP therapeutics and highlights recent advances in separation science for reliable characterization and quality control. We focus on techniques for RNA, LNPs, and the RNA-LNP complex, emphasizing chromatographic, electrophoretic, and field-based separation techniques.
基因疗法作为一种药物模式正在迅速发展，是治疗以前难以成药的途径的一种手段。在基于成簇规律间隔短回文重复序列（CRISPR）的基因编辑中使用小干扰核糖核酸（siRNA）、信使核糖核酸（mRNA）和引导核糖核酸（gRNA），并结合脂质纳米颗粒（LNP），已证明可实现有效的药物递送。这些复杂的递送系统通常需要多种分析方法来实现全面表征，其中一些方法仍未充分发展或不适用于RNA-LNP制剂。常用的基于批次的方法，如动态光散射（DLS）或改良的RiboGreen测定法，经常受到样品异质性的阻碍，这一限制可通过分析分离来解决。本综述讨论了限制RNA-LNP治疗药物分析分离的挑战，并强调了分离科学在可靠表征和质量控制方面的最新进展。我们重点介绍了用于RNA、LNP和RNA-LNP复合物的技术，强调了色谱、电泳和基于场的分离技术。
