Size-exclusion chromatography with post-column nucleic acid staining and fluorescence detection for sensitive ribonucleic acid analysis.

ABSTRACT: RNA-based therapeutics, such as vaccines or gene therapy applications, are steadily gaining in popularity and require extensive analytical characterization with several orthogonal methods to ensure highest purity for safe use. In this study, size-exclusion chromatography (SEC) with multiple detectors was used for characterizing single- and double-stranded (ss/ds) RNA ladders, providing insights into how conformation and viscosity affect the different elution profiles observed for both species in SEC. Furthermore, high-resolution chromatographic separation was merged with the high-sensitivity detection provided by fluorescent dyes by developing an innovative approach relying on SEC with post-column nucleic acid staining and fluorescence detection (SEC-pcs-FLD). Herein, SEC-pcs-FLD was succe

SUBMITTER: Hoevelmann Y 

PROVIDER: S-EPMC12325474 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature