mRNA Characterization, from 5’ Cap to Poly (A): What IP-RP-MS Can Tell You

Ms. Roxana Eggleston-Rangel, Advanced Workflows Applications Manager at Phenomenex

mRNA Characterization, from 5’ Cap to Poly (A): What IP-RP-MS Can Tell You

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Roxana Eggleston-Rangel received her bachelor’s and master’s degree in Chemistry and Biochemistry from California State University, Los Angeles. Roxana joined Phenomenex in 2018 as an Applications Scientist and is currently working with the applications team to produce analytical solutions for scientist in the biopharma industry. Roxana’s current work focuses on characterization of mRNA, sgRNA and siRNA. Prior to Phenomenex, Roxana worked for Caltech and the University of Southern California developing LM-CS workflows for the proteomics community.

mRNA is a biotherapeutic modality that has been successfully used in vaccines and vaccine development. Because of the fast-tracked success, high demand for robust analytical methods used to make critical quality decisions are needed. The manufacturing process of mRNA molecules consists of in vitro transcription reactions in which the desired final product is over 1000 nucleotides in length, has a 5’ Cap and a Poly (A) tail. Both the 5’ Cap and the Poly (A) tail help to stabilize the RNA molecule and improve its translation. Contaminants, such as degradation products (incomplete capping and shorter than expected Poly(A) tails), can still be present in the final product and need to be quantified to assess mRNA purity. This study shows an LC-MS characterization method for three mRNA key critical quality attributes: 5’ Cap, Open Reading Frame (ORF) and Poly (A) tail.