Background
Large Ribonucleic Acids (RNAs) including messenger RNAs (mRNAs) are promising candidates for therapeutic treatment and vaccines, having generated great interest in the field of vaccination as a safe and efficient alternative to traditional live virus or protein-based vaccines. Unlike traditional vaccines, mRNA can be engineered to carry specific genetic information, which upon delivery into cells and using host cell translational machinery, can result in generation of a desired antigen in vivo. While the process relies on the biological machinery naturally contained in the body, mRNA is susceptible to degradation during preparation, processing, formulation and long-term storage conditions. Furthermore, once introduced into the host, it is subject to many biological processes that can degrade it upon delivery.
A high-resolution, sensitive analytical method that can measure the integrity of mRNA molecules in terms of their size and length, is therefore crucial for quality assurance, understanding potency, and optimization of manufacturing processes that must be closely regulated and monitored throughout the whole manufacturing process and product release to ensure the final products are safe, effective, and high quality considering its mode of action1. Critical Quality Attributes (CQAs) which have been identified that dictate the performance of a mRNA construct to efficiently express the gene of interest are: 5’capping efficiency and structure, UTR structure (length and regulatory elements), Coding sequence, PolyA tail properties and mRNA purity (and integrity)2 (Figure 1). mRNA Purity, which is a crucial determinant of yield, can be impacted by various impurities like smaller oligonucleotides, because of abortive initiation events and dsRNA generated by self-complementary 3’extension. These impurities can generate different immunological responses when injected into humans. This white paper describes the Capillary Gel Electrophoresis (CGE) method which can be used to assess the purity/integrity of mRNA-based drug products (DP) and is accepted by all the regulatory agencies.
To continue reading, please fill out the form.