Analyzing Poly(A) Tails Of In Vitro Transcribed RNA With The Agilent Fragment Analyzer Systems

By Lisa Houston and Whitney Pike, Agilent Technologies

The synthesis of in vitro transcribed (IVT) mRNA requires the addition of a poly-A tail, a critical modification that stabilizes the molecule and promotes efficient translation. Accurately characterizing the length and quality of this poly-A tail is essential for ensuring the integrity and efficacy of mRNA drug substances and products.

This application note demonstrates a robust method for analyzing poly-A tails using automated capillary electrophoresis (CE). The method successfully separates the IVT mRNA from the free poly-A tail, allowing for the independent quantification of each component. Key results show that the CE system provides reliable and reproducible sizing and concentration data for both the main product and the poly-A tail. This technique offers a precise and high-throughput alternative to traditional gel-based methods, supporting faster and more accurate quality control during the development and manufacture of mRNA therapeutics.

Access the full application note to review the complete methodology and data for poly-A tail analysis.