Most researchers have taken mRNA as a tough material to work with For the ubiquitous presence of RNases that rapidly degrade RNA.
To avoid mRNA degradation, the manufacturing process, including in vitro transcription, adding 5’ cap and 3’ poly(A)-tail, and mRNA purification, has to be conducted with strictly RNase-free materials, hence the extensive testing of RNase contamination of all components and equipment before use. Standardization of the process contributes in maintaining batch-to-batch reproducibility.
The IVT mRNA sequence varies but its manufacturing process does not. After the production phase, mRNA products need to go through extensive testing and characterization to ensure the good quality, appearance, content, integrity, absence of contamination and the potential for mRNA to be translated into proteins.
We keep working on new analysis and test methods for mRNA quality control. Capillary electrophoresis, HPLC, and LC-Mass are used for the purity and integrity of mature mRNA, as well as the efficiency of capping and tailing.