用于快速灵敏检测体外转录mRNA产物中双链RNA污染物的侧向流动免疫分析
Lateral Flow Immunoassay for Rapid and Sensitive Detection of dsRNA Contaminants in In Vitro-Transcribed mRNA Products.
作者信息
Luo Dengwang, Wang Daming, Cao Yuhong
机构信息
State Key Laboratory of Synthetic Biology, Frontiers Science Center for Synthetic Biology (Ministry of Education), School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin, China.
CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Chinese Academy of Sciences, Beijing, China.
出版信息
Methods Mol Biol. 2025;2965:189-200. doi: 10.1007/978-1-0716-4742-4_7.
Double-stranded RNA (dsRNA), often generated as a by-product during in vitro transcription, poses a significant challenge to therapeutic applications of mRNA. In this chapter, we present a rapid and sensitive lateral flow strip assay (LFSA) utilizing colloidal gold nanoparticles for detecting dsRNA contaminants in in vitro-transcribed mRNA. The assay delivers results within 15 min and achieves a limit of detection (LOD) of 69.32 ng/mL for N1-methyl-pseudouridine-containing dsRNA. This method provides a quick and efficient tool for ensuring mRNA purity, reducing the risk of dsRNA-induced immunogenicity, and facilitating large-scale mRNA production without the need for time-consuming traditional methods.
双链RNA（dsRNA）通常是体外转录过程中产生的副产物，对mRNA的治疗应用构成重大挑战。在本章中，我们介绍了一种快速灵敏的侧向流动试纸条检测法（LFSA），该方法利用胶体金纳米颗粒检测体外转录mRNA中的dsRNA污染物。该检测法在15分钟内即可得出结果，对于含N1-甲基假尿苷的dsRNA，检测限（LOD）达到69.32 ng/mL。该方法为确保mRNA纯度、降低dsRNA诱导的免疫原性风险以及促进大规模mRNA生产提供了一种快速有效的工具，无需耗时的传统方法。
Zotero 插件