Residual dsRNA testing by ELISA in mRNA vaccines 

The presence of residual double-stranded RNA (dsRNA) in mRNA vaccines is a critical quality attribute with potential implications for product potency and immunogenicity. As a byproduct of in vitro transcription (IVT), dsRNA can trigger innate immune responses, leading to increased reactogenicity and reduced protein translation. Regulatory agencies, including The United States Pharmacopeia (USP), recognize the importance of dsRNA quantification and recommend validated analytical methods such as immunoblot and enzyme-linked immunosorbent assay (ELISA) for systematic release testing.

This white paper explores the development and optimization of an in-house ELISA for dsRNA detection, addressing key technical challenges such as antibody selection, assay calibration, matrix interference, and reagent variability. By leveraging a deep understanding of dsRNA structure and detection methodologies, we outline a robust approach to enhance the specificity, sensitivity, and reproducibility of dsRNA testing in mRNA vaccine development.