mRNA is produced by in vitro transcription (IVT) reaction, which is typically performed as a batch process. Considering its catalytic basis, it is possible to extend reaction time and yields by continuous addition of consumed reagents to the reaction mixture, however fed-batch strategies reported to date have only managed to achieve a 40 – 100 % increase in the production of mRNA. One of the main limitations of development has been low throughput of analytics for quantification of mRNA and NTP precursors; productivity is typically determined as an end-point measurement of mRNA concentration.
Webinar will discuss an HPLC-based analytical methodology using a multimodal ligand based on anion exchange/hydrogen-bonding ligand (PrimaS) to monitor the IVT reaction, which allows for simultaneous quantification of NTPs, capping reagent, plasmid, and mRNA within 3.5 min.
When applied to monitoring IVT reaction a batch approach with an average productivity of 3-5 mg/mL can be converted to fed-batch yielding 10-12 mg/mL. Two approaches for controlling IVT reaction will be demonstrated, one focusing on productivity of uncapped mRNA, another on productivity of capped mRNA, thereby requiring a precise control of NPT:capping reagent ratio
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