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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.

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Lateral Flow Immunoassay for Rapid and Sensitive Detection of dsRNA Contaminants in In Vitro-Transcribed mRNA Products

  • 实验方案

  • 发表日期：

    2025年

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  • Dengwang Luo,
  • Daming Wang,
  • Yuhong Cao

摘要

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.